2-anilino-4-aminoalkyleneaminopyrimidines

ABSTRACT

The present invention relates to 2-arylamino-4-(aminoalkylene)aminopyrimidines inhibitors having the structure shown below: 
                         
wherein R and R 1  are as defined herein. The 2-arylamino-4 -(aminoalkylene)aminopyrimidines inhibit Protein Kinase C-alpha (PKC-α). The PKC-α inhibitors of the present invention are important for improving myocardial intracellular calcium cycling, resulting in improved myocardial contraction and relaxation performance and thereby slowing the progression of heart failure. The present invention further relates to compositions comprising said 2-arylamino-4 -(aminoalkylene)amino-pyrimidines and to methods for controlling, abating, or otherwise slowing the progression of heart failure.

RELATED CASES

This application is a continuation of U.S. application Ser. No.11/762,406 filed Jun. 13, 2007, which claims priority benefit to U.S.provisional Application No. 60/813,875 filed Jun. 15, 2006.

FIELD OF THE INVENTION

The present invention relates to2-arylamino-4-(aminoalkylene)aminopyrimidines which are inhibitors ofProtein Kinase C-alpha (PKC-α). The PKC-α inhibitors of the presentinvention are important for improving myocardial intracellular calciumcycling, resulting in improved myocardial contraction and relaxationperformance and thereby slowing the progression of heart failure. Thepresent invention further relates to compositions comprising said2-arylamino-4-(aminoalkylene)amino-pyrimidines and to methods forcontrolling, abating, or otherwise slowing the progression of heartfailure.

BACKGROUND OF THE INVENTION

Many biologically active substances, for example, hormones,neurotransmitters and peptides are known to exert functions viaintracellular mediators such as, cyclic adenosine monophosphate (cAMP),cyclic guanosine monophosphate (cGMP), diacylglycerol (DAG) and calcium.In many cases, these mediators activate or inactivate intracellularkinases or phosphatases that are important in proteinphosphorylation/dephosphorylation, and thus play important roles inregulating cellular processes and functions. The protein kinase C (PKC)family of calcium and/or lipid-activated serine-threonine kinasesfunction downstream of nearly all membrane-associated signaltransduction pathways.¹ Approximately 12 different isozymes comprise thePKC family, which are broadly classified by their activationcharacteristics. The conventional PKC isozymes (PKCα, βI, βII, and γ)are calcium- and lipid-activated, while the novel isozymes (ε, θ, η, andδ) and atypical isozymes (ζ, ι, ν, and μ) are calcium independent butactivated by distinct lipids.² For example, stimulation of Gαq-coupledG-protein coupled receptors (GPCR) can activate phospholipase C (PLC)which in turn mediates hydrolysis of inositol phospholipids resulting inthe generation of inositol 1,4,5-triphosphate (IP₃) and DAG. IP₃ and DAGcan activate the different isoforms of PKC by mobilizing calcium(calcium sensitive enzymes) or by directly activating PKC, respectively.Once activated, PKC isozymes translocate to discrete subcellularlocations through direct interactions with docking proteins termed RACKs(Receptor for Activated C Kinases), which permit specific substraterecognition and subsequent signal transduction.³

Alterations in PKC activity has been suggested to contribute to humandiseases, inter alia, diabetes, numerous forms of cancer,microalbinuria, endothelial dysfunction, cerebrovascular disease,stroke, coronary heart disease, cardiovascular disease and sequela (e.g.arrhythmia, sudden death, increased infarct size, congestive heartfailure, angina), myocardial ischemic states, hypertension, lipiddisorders, ischemia-reperfusion injury, atherosclerosis, peripheralartery/vascular disease, microvascular complications of diabetes(neuropathy, nephropathy, retinopathy), restenosis, renal disease, bloodcoagulation disorders, inflammatory diseases and heart failure andinhibition of PKC in these settings could be used to treat or preventhuman disease. Lending support to the modulation of PKC in cardiacdisease, PKC activation has been associated with cardiac hypertrophy,dilated cardiomyopathy, ischemic injury and mitogen stimulation.

Heart disease is the leading cause of death in industrialized nations.Historically heart failure (HF) has been a product of hypertension,coronary heart disease, genetic disorders, valvular deformities,diabetes or cardiomyopathy. While the root cause of heart failure ismultifaceted, it uniformly is marked by impaired diastolic and/orsystolic function and can be accompanied by chamber enlargement whichultimately manifest in symptomatic heart failure (fatigue, pulmonaryedema, circulatory congestion, etc.)

The risk of death due to heart failure is 5-10% annually in patientswith mild symptoms of heart failure, and increases to 30-40% annually inpatients with advanced heart failure, with a 50% overall mortality rateat 5 years. The current mainstays of heart failure therapy are drugsthat act on the renin-angiotensin-aldosterone system (ACEI, ARB,aldosterone inhibitor), diuretics, digoxin and β-adrenergic receptorblockers. Despite the fact that multiple drug classes are used to treatheart failure patients, new cases of heart failure are growing at over10% per year.

Patients with acute decompensated heart failure (ADHF) are a treatmentchallenge to physicians and can present with volume overload and/ordiminished cardiac output. Initial treatments for ADHF patients includeintravenous diuretics, vasodilators, natriuretic peptides and inotropicagents. Despite the widespread use of these agents, long-term safety andbenefit of these drugs have been questioned. In the case of inotropes,drugs that increases cardiac output and cardiac contractility withoutincreasing myocardial oxygen consumption or heart rate are desirous.Despite the available treatments for patients with ADHF, hospitalreadmission rates are approximately 50% within 6 months and mortality isapproximately 20-40% at 1 year.

The primary function of the heart is to generate and sustain an arterialblood pressure necessary to provide adequate perfusion of organs. Ithas, therefore, become an area of intense investigation to decipher themechanism(s) which initiate and contribute to the development of heartfailure rather than relying on a means for treating the symptoms ofheart failure alone. At the cardiomyocyte (cardiac contractile cells)level, impaired calcium cycling is a hallmark of heart failure as is thebasis of contractile abnormalities. Calcium plays a key role inregulating kinases, phosphatases and transcription factors believed toinfluence the remodeling process indicating that both acute andsustained alterations in intracellular calcium levels may have profoundeffect on cardiac function and remodeling (i.e., changes in wallthickness or chamber volume). This theory would support the propositionthat the development of new therapies addressing the slowing andpreventing of the disease progression, would be perhaps more effectiveagainst heart failure than palliation of heart failure.

Therefore, there is a limited means to treat patients with various formsand stages of heart failure and there is incentive to develop novel,safe and effective treatments to prevent or treat patients with symptomsof heart failure, acute exacerbation of heart failure and chronic heartfailure and other cardiovascular diseases. An agent that has benefits inthe treating acute exacerbations of heart failure as well as treatingchronic heart failure is desirous.

-   1. Molkentin et al. (2001) Annu. Rev. Physiol. 63:391-426.-   2. Dempsey et al. (2000) Am. J. Physiol. Lung Mol. Physiol.    279:247-251.-   3. Mochly-Rosen, D. (1995) Science 268:247-251.

SUMMARY OF THE INVENTION

The present invention meets the aforementioned needs in that it has beenfound that certain 2-arylamino-4-(aminoalkylene)aminopyrimidines areeffective for inhibiting Protein Kinase C-alpha (PKC-α) therebyimproving myocardial contraction and relaxation performance and slowingthe progression of heart failure.

The present invention encompasses three major aspects each of which havetheir own separate categories, aspects, iterations, and specificiterative examples. The major aspects of the present invention include:

-   -   i) novel compositions of matter which are effective in        inhibiting PKC-α;    -   ii) compositions or pharmaceutical compositions (matrices)        comprising said compositions of matter;    -   iii) methods for controlling, abating, or alleviating one or        more of the causes of progressive heart failure which is        affected by administration of a PKC-α inhibitors, whether        administered alone or in a composition or within a        pharmaceutical composition (matrix); and    -   iv) a process for preparing the PKC-α inhibitors of the present        invention.

The first aspect of the present invention as a whole, relates tocompounds, which include all enantiomeric and diastereomeric forms andpharmaceutically acceptable salts thereof, said compounds having theformula:

wherein R is a unit having the formula:

R² and R³ are each independently chosen from:

-   -   i) hydrogen; or    -   ii) C₁-C₄ substituted or unsubstituted linear, branched, or        cyclic alkyl;

L is a linking unit having the formula:—[C(R^(4a)R^(4b))]_(n)—

each R⁴ is independently chosen from

-   -   i) hydrogen; or    -   ii) C₁-C₄ alkyl;

the index n is from 1 to 4; and

R¹ is substituted or unsubstituted aryl.

The second major aspect of the present invention relates to compositionscomprising:

-   -   a) an effective amount of one or more compounds according to the        present invention; and    -   b) one or more acceptable excipients.

The third major aspect of the present invention relates to methods ofuse. As described herein below, the PKC-α inhibitors of the presentinvention are important for improving myocardial contraction andrelaxation performance and thereby slowing the progression of heartfailure and their administration to humans is, therefore, an effectivetreatment for humans suffering from acute heart failure.

These and other objects, features, and advantages will become apparentto those of ordinary skill in the art from a reading of the followingdetailed description and the appended claims. All percentages, ratiosand proportions herein are by weight, unless otherwise specified. Alltemperatures are in degrees Celsius (° C.) unless otherwise specified.All documents cited are in relevant part, incorporated herein byreference; the citation of any document is not to be construed as anadmission that it is prior art with respect to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention addresses several unmet medical needs, inter alia:

-   -   1) improving cardiac contraction/relaxation parameters in heart        failure patients, leading to reduction of symptoms; and    -   2) attenuating adverse cardiac remodeling in heart failure        patients, ultimately providing prolonged patient survival.

These and other unmet medical needs are resolved by the PKC-α inhibitorsof the present invention, which are capable of blocking Protein KinaseC-alpha from impairing sarcoplasmic reticulum Ca²⁺ uptake. By providingheart failure patients with a PKC-α inhibitor, it is believed thepatients will derive an improvement in cardiac function, thus resultingin improved myocardial contraction and relaxation performance and couldresult in slowing the progression to heart failure.

The following chemical hierarchy is used throughout the specification todescribe and enable the scope of the present invention and toparticularly point out and distinctly claim the units which comprise thecompounds of the present invention. The term “hydrocarbyl” stands forany carbon atom-based unit (organic molecule), said units optionallycontaining one or more organic functional group, including inorganicatom comprising salts, inter alia, carboxylate salts, quaternaryammonium salts. Within the broad meaning of the term “hydrocarbyl” arethe classes “acyclic hydrocarbyl” and “cyclic hydrocarbyl” which termsare used to divide hydrocarbyl units into cyclic and non-cyclic classes.

As it relates to the following definitions, “cyclic hydrocarbyl” unitsmay comprise only carbon atoms in the ring (hydrocarbyl and aryl rings)or may comprise one or more heteroatoms in the ring (heterocyclic andheteroaryl). For “hydrocarbyl” rings the lowest number of carbon atomsin a ring are 3 carbon atoms; cyclopropyl. For “aryl” rings the lowestnumber of carbon atoms in a ring are 6 carbon atoms; phenyl. For“heterocyclic” rings the lowest number of carbon atoms in a ring is 1carbon atom; diazirinyl, epoxy. For “heteroaryl” rings the lowest numberof carbon atoms in a ring is 1 carbon atom; 1,2,3,4-tetrazolyl. Thefollowing is a non-limiting description of the terms “acyclichydrocarbyl” and “cyclic hydrocarbyl” as used herein.

A. Substituted and Unsubstituted C₁-C₂₀ Acyclic Hydrocarbyl:

-   -   For the purposes of the present invention the term “substituted        and unsubstituted C₁-C₂₀ acyclic hydrocarbyl” encompasses 3        categories of units:

-   1) C₁-C₂₀ linear or branched alkyl, non-limiting examples of which    include, methyl (C₁), ethyl (C₂), n-propyl (C₃), iso-propyl (C₃),    n-butyl (C₄), sec-butyl (C₄), iso-butyl (C₄), tert-butyl (C₄), and    the like; substituted C₁-C₂₀ linear or branched alkyl, non-limiting    examples of which includes, hydroxymethyl (C₁), chloromethyl (C₁),    trifluoromethyl (C₁), aminomethyl (C₁), 1-chloroethyl (C₂),    2-hydroxyethyl (C₂), 1,2-difluoroethyl (C₂), 3-carboxypropyl (C₃),    and the like.

-   2) C₂-C₂₀ linear or branched alkenyl, non-limiting examples of which    include, ethenyl (C₂), 3-propenyl (C₃), 1-propenyl (also    2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),    buten-4-yl (C₄), and the like; substituted C₂-C₂₀ linear or branched    alkenyl, non-limiting examples of which include, 2-chloroethenyl    (also 2-chlorovinyl) (C₂), 4-hydroxybuten-1-yl (C₄),    7-hydroxy-7-methyloct-4-en-2-yl (C₉),    7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like.

-   3) C₂-C₂₀ linear or branched alkynyl, non-limiting examples of which    include, ethynyl (C₂), prop-2-ynyl (also propargyl) (C₃),    propyn-1-yl (C₃), and 2-methyl-hex-4-yn-1-yl (C₇); substituted    C₂-C₂₀ linear or branched alkynyl, non-limiting examples of which    include, 5-hydroxy-5-methylhex-3-ynyl (C₇),    6-hydroxy-6-methylhept-3-yn-2-yl (C₈), 5-hydroxy-5-ethylhept-3-ynyl    (C₉), and the like.    B. Substituted and Unsubstituted C₁-C₂₀ Cyclic Hydrocarbyl:    -   For the purposes of the present invention the term “substituted        and unsubstituted C₁-C₂₀ cyclic hydrocarbyl” encompasses 5        categories of units:

-   1) The term “carbocyclic” is defined herein as “encompassing rings    comprising from 3 to 20 carbon atoms, wherein the atoms which    comprise said rings are limited to carbon atoms, and further each    ring can be independently substituted with one or more moieties    capable of replacing one or more hydrogen atoms.” The following are    non-limiting examples of “substituted and unsubstituted C₃-C₂₀    carbocyclic rings” which encompass the following categories of    units:    -   i) carbocyclic rings having a single substituted or        unsubstituted hydrocarbon ring, non-limiting examples of which        include, cyclopropyl (C₃), 2-methyl-cyclopropyl (C₃),        cyclopropenyl (C₃), cyclobutyl (C₄), 2,3-dihydroxycyclobutyl        (C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),        cyclopentadienyl (C₅), cyclohexyl (C₆), cyclohexenyl (C₆),        cycloheptyl (C₇), cyclooctanyl (C₈), decalinyl (C₁₀),        2,5-dimethylcyclopentyl (C₅), 3,5-dichlorocyclohexyl (C₆),        4-hydroxycyclohexyl (C₆), and 3,3,5-trimethylcyclohex-1-yl (C₆).    -   ii) carbocyclic rings having two or more substituted or        unsubstituted fused hydrocarbon rings, non-limiting examples of        which include, octahydropentalenyl (C₈), octahydro-1H-indenyl        (C₉), 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl (C₉),        decahydroazulenyl (C₁₀); bicyclo[6.2.0]decanyl (C₁₀),        decahydronaphthalenyl (C₁₀), and dodecahydro-1H-fluorenyl (C₁₃).    -   iii) carbocyclic rings which are substituted or unsubstituted        bicyclic hydrocarbon rings, non-limiting examples of which        include, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl,        bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl,        bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

-   2) The term “aryl” is defined herein as “units encompassing at least    one phenyl or naphthyl ring and wherein there are no heteroaryl or    heterocyclic rings fused to the phenyl or naphthyl ring and further    each ring can be independently substituted with one or more moieties    capable of replacing one or more hydrogen atoms.” The following are    non-limiting examples of “substituted and unsubstituted C₆-C₁₄ aryl    rings” which encompass the following categories of units:    -   i) C₆ or C₁₀ substituted or unsubstituted aryl rings; phenyl and        naphthyl rings whether substituted or unsubstituted,        non-limiting examples of which include, phenyl (C₆),        naphthylen-1-yl (C₁₀), naphthylen-2-yl (C₁₀), 4-fluorophenyl        (C₆), 2-hydroxyphenyl (C₆), 3-methylphenyl (C₆),        2-amino-4-fluorophenyl (C₆), 2-(N,N-diethylamino)phenyl (C₆),        2-cyanophenyl (C₆), 2,6-di-tert-butylphenyl (C₆),        3-methoxyphenyl (C₆), 8-hydroxynaphthylen-2-yl (C₁₀),        4,5-dimethoxynaphthylen-1-yl (C₁₀), and 6-cyano-naphthylen-1-yl        (C₁₀).    -   ii) C₆ or C₁₀ aryl rings fused with 1 or 2 saturated rings        non-limiting examples of which include,        bicyclo[4.2.0]octa-1,3,5-trienyl (C₈), and indanyl (C₉).

-   3) The terms “heterocyclic” and/or “heterocycle” are defined herein    as “units comprising one or more C₁-C₂₀ rings having from 3 to 20    atoms wherein at least one atom in at least one ring is a heteroatom    chosen from nitrogen (N), oxygen (O), or sulfur (S), or mixtures of    N, O, and S, and wherein further the ring which comprises the    heteroatom is also not an aromatic ring.” The following are    non-limiting examples of “substituted and unsubstituted C₁-C₂₀    heterocyclic rings” which encompass the following categories of    units:    -   i) heterocyclic units having a single ring containing one or        more heteroatoms, non-limiting examples of which include,        diazirinyl (C₁), aziridinyl (C₂), urazolyl (C₂), azetidinyl        (C₃), pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl        (C₃), isoxazolinyl (C₃), isoxazolyl (C₃), thiazolidinyl (C₃),        isothiazolyl (C₃), isothiazolinyl (C₃), oxathiazolidinonyl (C₃),        oxazolidinonyl (C₃), hydantoinyl (C₃), tetrahydrofuranyl (C₄),        pyrrolidinyl (C₄), morpholinyl (C₄), piperazinyl (C₄),        piperidinyl (C₄), dihydropyranyl (C₅), tetrahydropyranyl (C₅),        piperidin-2-onyl (valerolactam) (C₅),        2,3,4,5-tetrahydro-1H-azepinyl (C₆), 2,3-dihydro-1H-indole (C₈),        and 1,2,3,4-tetrahydro-quinoline (C₉).    -   ii) heterocyclic units having 2 or more rings one of which is a        heterocyclic ring, non-limiting examples of which include        hexahydro-1H-pyrrolizinyl (C₇),        3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl (C₇),        3a,4,5,6,7,7a-hexahydro-1H-indolyl (C₈),        1,2,3,4-tetrahydroquinolinyl (C₉), and        decahydro-1H-cycloocta[b]pyrrolyl (C₁₀).

-   4) The term “heteroaryl” is defined herein as “encompassing one or    more C₁-C₂₀ rings comprising from 5 to 20 atoms wherein at least one    atom in at least one ring is a heteroatom chosen from nitrogen (N),    oxygen (O), or sulfur (S), or mixtures of N, O, and S, and wherein    further at least one of the rings which comprises a heteroatom is an    aromatic ring.” The following are non-limiting examples of    “substituted and unsubstituted C₁-C₂₀ heterocyclic rings” which    encompass the following categories of units:    -   i) heteroaryl rings containing a single ring, non-limiting        examples of which include, 1,2,3,4-tetrazolyl (C₁),        [1,2,3]triazolyl (C₂), [1,2,4]triazolyl (C₂), triazinyl (C₃),        thiazolyl (C₃), 1H-imidazolyl (C₃), oxazolyl (C₃), furanyl (C₄),        thiopheneyl (C₄), pyrimidinyl (C₄), 2-phenylpyrimidinyl (C₄),        pyridinyl (C₅), 3-methylpyridinyl (C₅), and        4-dimethylaminopyridinyl (C₅).    -   ii) heteroaryl rings containing 2 or more fused rings one of        which is a heteroaryl ring, non-limiting examples of which        include: 7H-purinyl (C₅), 9H-purinyl (C₅), 6-amino-9H-purinyl        (C₅), 5H-pyrrolo[3,2-d]pyrimidinyl (C₆),        7H-pyrrolo[2,3-]pyrimidinyl (C₆), pyrido[2,3-]pyrimidinyl (C₇),        2-phenylbenzo[d]thiazolyl (C₇), 1H-indolyl (C₈),        4,5,6,7-tetrahydro-1-H-indolyl (C₈), quinoxalinyl (C₈),        5-methylquinoxalinyl (C₈), quinazolinyl (C₈), quinolinyl (C₉),        8-hydroxy-quinolinyl (C₉), and isoquinolinyl (C₉).

-   5) C₁-C₆ tethered cyclic hydrocarbyl units (whether C₃-C₁₀    carbocyclic units, C₆ or C₁₀ aryl units, C₁-C₁₀ heterocyclic units,    or C₁-C₁₀ heteroaryl units) which connected to another moiety, unit,    or core of the molecule by way of a C₁-C₆ alkylene unit.    Non-limiting examples of tethered cyclic hydrocarbyl units include    benzyl C₁-(C₆) having the formula:

-   -   wherein R^(a) is optionally one or more independently chosen        substitutions for hydrogen. Further examples include other aryl        units, inter alia, (2-hydroxyphenyl)hexyl C₆—(C₆);        naphthalen-2-ylmethyl C₁-(C₁₀), 4-fluorobenzyl C₁-(C₆),        2-(3-hydroxy-phenyl)ethyl C₂—(C₆), as well as substituted and        unsubstituted C₃-C₁₀ alkylenecarbocyclic units, for example,        cyclopropylmethyl C₁-(C₃), cyclopentylethyl C₂—(C₅),        cyclohexylmethyl C₁-(C₆). Included within this category are        substituted and unsubstituted C₁-C₁₀ alkylene-heteroaryl units,        for example a 2-picolyl C₁-(C₆) unit having the formula:

-   -   wherein R^(a) is the same as defined above. In addition, C₁-C₁₂        tethered cyclic hydrocarbyl units include C₁-C₁₀        alkyleneheterocyclic units and alkylene-heteroaryl units,        non-limiting examples of which include, aziridinylmethyl C₁-(C₂)        and oxazol-2-ylmethyl C₁-(C₃).

For the purposed of the present invention fused ring units, as well asspirocyclic rings, bicyclic rings and the like, which comprise a singleheteroatom will be considered to belong to the cyclic familycorresponding to the heteroatom containing ring. For example,1,2,3,4-tetrahydroquinoline having the formula:

is, for the purposes of the present invention, considered a heterocyclicunit. 6,7-Dihydro-5H-cyclopentapyrimidine having the formula:

is, for the purposes of the present invention, considered a heteroarylunit. When a fused ring unit contains heteroatoms in both a saturatedand an aryl ring, the aryl ring will predominate and determine the typeof category to which the ring is assigned. For example,1,2,3,4-tetrahydro-[1,8]naphthyridine having the formula:

is, for the purposes of the present invention, considered a heteroarylunit.

The term “substituted” is used throughout the specification. The term“substituted” is defined herein as “a hydrocarbyl moiety, whetheracyclic or cyclic, which has one or more hydrogen atoms replaced by asubstituent or several substituents as defined herein below.” The units,when substituting for hydrogen atoms are capable of replacing onehydrogen atom, two hydrogen atoms, or three hydrogen atoms of ahydrocarbyl moiety at a time. In addition, these substituents canreplace two hydrogen atoms on two adjacent carbons to form saidsubstituent, new moiety, or unit. For example, a substituted unit thatrequires a single hydrogen atom replacement includes halogen, hydroxyl,and the like. A two hydrogen atom replacement includes carbonyl,oximino, and the like. A two hydrogen atom replacement from adjacentcarbon atoms includes epoxy, and the like. Three hydrogen replacementincludes cyano, and the like. The term substituted is used throughoutthe present specification to indicate that a hydrocarbyl moiety, interalia, aromatic ring, alkyl chain; can have one or more of the hydrogenatoms replaced by a substituent. When a moiety is described as“substituted” any number of the hydrogen atoms may be replaced. Forexample, 4-hydroxyphenyl is a “substituted aromatic carbocyclic ring”,(N,N-dimethyl-5-amino)octanyl is a “substituted C₈ alkyl unit,3-guanidinopropyl is a “substituted C₃ alkyl unit,” and2-carboxypyridinyl is a “substituted heteroaryl unit.”

The following are non-limiting examples of categories and examplesherewith of units which can suitably substitute for hydrogen atoms on acyclic or acyclic hydrocarbyl unit, described herein below as R⁵ units,wherein in the non-limiting examples provided herein below, R⁶ ishydrogen, C₁-C₁₀ linear or branched alkyl, C₂-C₁₀ linear or branchedalkenyl, C₂-C₁₀ linear or branched alkynyl, and C₆ or C₁₀ aryl.

R⁵ units according to the present invention include the following:

-   -   i) —NHCOR⁶; for example, —NHCOCH₃, —NHCOCH₂CH₃, —NHCOC₆H₅;    -   ii) —COR⁶; for example, —COCH₃, —COCH₂CH₃, —COCH₂CH₂CH₃;    -   iii) —CO₂R⁶; for example, —CO₂CH₃, —CO₂CH₂CH₃, —CO₂CH₂CH₂CH₃;    -   iv) —OCOR⁶; for example, —OCOCH₃, —OCOCH₂CH₃, —OCOCH₂CH₂CH₃;    -   v) —C(═NH)NH₂;    -   vi) —NHC(═NH)NH₂;    -   vii) —N(R⁶)₂; for example, —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃);    -   viii) —NHC₆H₅;    -   ix) C₁-C₄ linear, branched, or cyclic alkyl; for example,        methyl, ethyl;    -   x) —CON(R⁶)₂; for example, —CONH₂, —CONHCH₃, —CON(CH₃)₂;    -   xi) —CONHNH₂;    -   xii) —NHCN;    -   xiii) —CN;    -   xiv) halogen: —F, —Cl, —Br, and —I;    -   xv) —NHN(R⁶)₂; for example, —NHNH₂, —NHNHCH₃, —NHN(CH₃)₂;    -   xvi) —OR⁶; for example, —OH, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃;    -   xvii) —NO₂;    -   xviii) —CH_(m)X_(n); wherein X is halogen, m is from 0 to 2,        m+n=3; for example, —CH₂F, —CHF₂, —CF₃, —CCl₃, or —CBr₃; and    -   xix) —SO₂N(R⁶)₂; for example, —SO₂NH₂; —SO₂NHCH₃; —SO₂NHC₆H₅.

For the purposes of the present invention the terms “compound” and“analog” stand equally well for the novel compositions of matterdescribed herein, including all enantiomeric forms, diastereomericforms, salts, and the like, and the terms “compound” and “analog” areused interchangeably throughout the present specification.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of suitable acids includehydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric,maleic, phosphoric, glycolic, lactic, salicylic, succinic,toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic,benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acids.Other acids, such as oxalic acid, while not themselves pharmaceuticallyacceptable, may be employed in the preparation of salts useful asintermediates in obtaining the compounds of this invention and theirpharmaceutically acceptable acid addition salts. Salts derived fromappropriate bases include alkali metal (e.g., sodium), alkaline earthmetal (e.g., magnesium), ammonium and N—(C₁-C₄ alkyl)₄+salts.

The compounds of the present invention are2-arylamino-4-(aminoalkylene)-aminopyrimidines having the core scaffold:

R units are amino units having the formula:

wherein R² and R³ are each independently chosen from:

-   -   i) hydrogen; or    -   ii) C₁-C₄ substituted or unsubstituted linear, branched, or        cyclic alkyl.

The first aspect of R units relates to amino units wherein R₂ and R₃ areindependently chosen from hydrogen, methyl (C₁), or ethyl (C₂), saidunits having the formula:

-   -   i) —NH₂;    -   ii) —NHCH₃;    -   iii) —N(CH₃)₂;    -   iv) —NH(CH₂CH₃);    -   v) —N(CH₂CH₃)₂; and    -   vi) —N(CH₃)(CH₂CH₃).

The second aspect of R units relates to amino units wherein R² and R³are independently chosen from hydrogen, n-propyl (C₃), or iso-propyl(C₃), said units having the formula:

-   -   i) —NH(CH₂CH₂CH₃);    -   ii) —N(CH₂CH₂CH₃)₂;    -   iii) —NH[CH(CH₃)₂];    -   iv) —N[CH(CH₃)₂]₂; and    -   v) —N(CH₂CH₂CH₃)[CH(CH₃)₂].    -   vi)

The third aspect of R units relates to amino units wherein R² is chosenfrom methyl (C₁) or ethyl (C₂), and R³ is chosen from n-propyl (C₃) oriso-propyl (C₃), said units having the formula:

-   -   i) —N(CH₃)(CH₂CH₂CH₃);    -   ii) —N(CH₂CH₃)(CH₂CH₂CH₃);    -   iii) —N(CH₃)[CH(CH₃)₂]; and    -   iv) —N(CH₂CH₃)[CH(CH₃)₂]₂.    -   v)    -   The fourth aspect of R units relates to amino units wherein R²        and R³ are independently chosen from hydrogen, n-butyl (C₄),        iso-butyl (C₄), sec-butyl (C₄), and tert-butyl (C₄),        non-limiting examples of said units having the formula:    -   i) —NH(CH₂CH₂CH₂CH₃);    -   ii) —N(CH₂CH₂CH₂CH₃)₂;    -   iii) —NH[CH₂CH(CH₃)₂];    -   iv) —N[CH₂CH(CH₃)₂]₂;    -   v) —NH[CH(CH₃)CH₂CH₃];    -   vi) —N[CH(CH₃)CH₂CH₃]₂;    -   vii) —NH[C(CH₃)₃];    -   viii) —N[C(CH₃)₃]₂    -   ix) —N(CH₂CH₂CH₂CH₃)[CH₂CH(CH₃)₂];    -   x) —N(CH₂CH₂CH₂CH₃)[CH(CH₃)CH₂CH₃]; and    -   xi) —N(CH₂CH₂CH₂CH₃)[C(CH₃)₃].

R¹ is substituted or unsubstituted aryl having the formula:

wherein R⁵ represents one or more (from 1 to 5) optionally present, andindependently selected, substitutes for hydrogen as outlined hereinabove and described in the categories, aspects, iterations, examples,and tables herein below.

The first category of R¹ units relates to aryl units which aresubstituted by one or more R⁵ units chosen from:

-   -   i) C₁-C₄ linear, branched, or cyclic alkyl; for example, methyl        (C₁), ethyl (C₂), n-propyl (C₃), isopropyl (C₃), n-butyl (C₄),        and the like;    -   ii) halogen; for example, —F, —Cl, —Br, and —I;    -   iii) —OR⁶; for example, —OCH₃ (C₁), —OCH₂CH₃ (C₂), —OCH₂CH₂CH₃        (C₃), —OCH(CH₃)₂ (C₃), and the like;    -   iv) —SO₂N(R⁶)₂; for example, —SO₂NH₂, —SO₂NHCH₃, —SO₂NHC₆H₅, and        the like;    -   v) —CH_(m)X_(n); wherein each X is independently F, Cl, Br, or        I, m is from 0 to 2, m+n=3; for example, —CH₂F, —CHF₂, —CF₃,        —CCl₃, —CBr₃, and the like; and    -   vi) —NO₂;        wherein each R⁶ is independently hydrogen, or C₁-C₄ linear,        branched, or cyclic alkyl.

The first aspect of the first category of R¹ units relates tosubstituted phenyl units chosen from: 3-chlorophenyl, 4-chlorophenyl,3,4-dichlorophenyl, 3-chloro-4-methyl-phenyl, 3-chloro-4-fluorophenyl,3,4-difluorophenyl, 3-trifluoromethylphenyl,3-trifluoro-methyl-4-chlorophenyl, 3-methoxyphenyl, 3-methylphenyl,3-ethylphenyl, and 3-iso-propylphenyl.

The second aspect of the first category of R¹ units relates tosubstituted phenyl units chosen from: 2-fluorophenyl, 3-fluorophenyl,4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl,2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4-trifluorophenyl,2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl,2,4,6-trifluorophenyl, 2-chlorophenyl, 2,3-dichlorophenyl,2,4-dichlorophenyl, 2,5-dichlorophenyl, 2,6-dichloro-phenyl,2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl, 2,3,6-trichlorophenyl,2,4,5-tri-chlorophenyl, and 2,4,6-trichlorophenyl.

The third aspect of the first category of R¹ units relates tosubstituted phenyl units chosen from: 2-methylphenyl, 4-methylphenyl,2,3-dimethylphenyl, 2,4-dimethylphenyl, 2,5-dimethyl-phenyl,2,6-dimethylphenyl, 3,4-dimethyl-phenyl, 2,3,4-trimethylphenyl,2,3,5-trimethyl-phenyl, 2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl,2,4,6-trimethyl-phenyl, 2-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl,2,4-diethylphenyl, 2,5-diethyl-phenyl, 2,6-diethylphenyl,3,4-diethylphenyl, 2,3,4-triethylphenyl, 2,3,5-triethylphenyl,2,3,6-triethylphenyl, 2,4,5-triethylphenyl, and 2,4,6-triethylphenyl.

The fourth aspect of the first category of R¹ units relates tosubstituted phenyl units chosen from: 2-methoxyphenyl, 4-methoxyphenyl,2,3-dimethoxyphenyl, 2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl,2,6-dimethoxyphenyl, 3,4-dimethoxyphenyl, 2,3,4-trimethoxyphenyl,2,3,5-trimethoxyphenyl, 2,3,6-trimethoxy-phenyl,2,4,5-tri-methoxyphenyl, 2,4,6-trimethoxyphenyl, 2-hydroxyphenyl,3-hydroxyphenyl, 4-hydroxy-phenyl, 2,3-dihydroxyphenyl,2,4-dihydroxyphenyl, 2,5-dihydroxyphenyl, 2,6-dihydroxy-phenyl,3,4-dihydroxy-phenyl, 2,3,4-trihydroxyphenyl, 2,3,5-trihydroxy-phenyl,2,3,6-trihydroxyphenyl, 2,4,5-trihydroxyphenyl, and2,4,6-trihydroxyphenyl.

However, other combinations of R⁵ units not specifically exemplified areencompassed within the first category of R¹.

The second category of R¹ units relates to aryl units which aresubstituted by one or more R⁵ units which are substituted orunsubstituted C₆ (phenyl) or C₁₀ (naphthyl) aryl units.

The first aspect of this category relates to R¹ units which are biphenylwherein R⁵ is phenyl, for example, a compound having the formula:

The second aspect of this category relates to R¹ units which aresubstituted biphenyl non-limiting examples of which include2′-fluoro-biphenyl-2-yl, 2′-fluoro-biphenyl-3-yl,2′-fluoro-biphenyl-4-yl, 3′-fluoro-biphenyl-2-yl,3′-fluoro-biphenyl-3-yl, 3′-fluoro-biphenyl-4-yl,4′-fluoro-biphenyl-2-yl, 4′-fluoro-biphenyl-3-yl,4′-fluoro-biphenyl-4-yl, 2′-chloro-biphenyl-2-yl,2′-chloro-biphenyl-3-yl, 2′-chloro-biphenyl-4-yl,3′-chloro-biphenyl-2-yl, 3′-chloro-biphenyl-3-yl,3′-chloro-biphenyl-4-yl, 4′-chloro-biphenyl-2-yl,4′-chloro-biphenyl-3-yl, 4′-chloro-biphenyl-4-yl,2′-methyl-biphenyl-2-yl, 2′-methyl-biphenyl-3-yl,2′-methyl-biphenyl-4-yl, 3′-methyl-biphenyl-2-yl,3′-methyl-biphenyl-3-yl, 3′-methyl-biphenyl-4-yl,4′-methyl-biphenyl-2-yl, 4′-methyl-biphenyl-3-yl, and4′-methyl-biphenyl-4-yl.

However, other combinations of R⁵ units not specifically exemplified areencompassed within the second category of R¹.

The third category of R¹ units relates to phenyl units which aresubstituted by one or more R⁵ units which are substituted orunsubstituted C₃, C₄ or C₅ heteroaryl units.

The first aspect of this category relates to R¹ units which are C₄ or C₅heteroaryl substituted phenyl units wherein R⁵ is, for example,pyridin-2-yl thereby forming a compound having the formula:

Non-limiting examples or this category of R¹ include2-pyridin-2-yl-phenyl, 3-pyridin-2-yl-phenyl, 4-pyridin-2-yl-phenyl,2-pyridin-3-yl-phenyl, 3-pyridin-3-yl-phenyl, 4-pyridin-3-yl-phenyl,2-pyridin-4-yl-phenyl, 3-pyridin-4-yl-phenyl, 4-pyridin-4-yl-phenyl,2-pyrimidin-2-yl-phenyl, 3-pyrimidin-2-yl-phenyl,4-pyrimidin-2-yl-phenyl, 2-pyrimidin-3-yl-phenyl,3-pyrimidin-3-yl-phenyl, 4-pyrimidin-3-yl-phenyl,2-pyrimidin-4-yl-phenyl, 3-pyrimidin-4-yl-phenyl, and4-pyrimidin-4-yl-phenyl.

The second aspect of this category relates to R¹ units which are C₃heteroaryl substituted phenyl wherein R⁵ is, for example,2-methyl-thiazol-4-yl thereby forming a compound having the formula:

Non-limiting examples of this aspect of the third category of R¹ includeimidazol-1-yl, imidazol-2-yl, imidazol-4-yl, thiazol-2-yl, thiazol-4-yl,and the like.

L is a linking unit having the formula:—[C(R^(4a)R^(4b))]_(n)—wherein each R^(4a) and R^(4b) unit is independently chosen from:

-   -   i) hydrogen; or    -   ii) C₁-C₄ linear, branched, or cyclic alkyl; for example, methyl        (C₁), ethyl (C₂), n-propyl(C₃), iso-propyl (C₃), cyclopropyl        (C₃), n-butyl (C₄), iso-butyl (C₄), sec-butyl (C₄), and        tert-butyl (C₄);        the index n is from 1 to 4. The index n indicates the number of        units which comprise L linking units, for example, a linking        unit having the formula —CH₂— (methylene) would have an index n        equal to 1. A linking unit having the formula —CH₂CH₂—        (ethylene) or the unit having the formula —CH(CH₃)CH₂—        (2-propylene) each have an index n equal to 2.

The first category of L units relates to unsubstituted alkylene unitschosen from:

-   -   i) —CH₂—, methylene;    -   ii) —CH₂CH₂—, ethylene;    -   iii) —CH₂CH₂CH₂—, propylene; and    -   iv) —CH₂CH₂CH₂CH₂—, butylene.

The first aspect of the first category of L units encompasses linkinggroups which are —CH₂CH₂CH₂—, propylene; as exemplified in the genericformula:

The second aspect of the first category of L units encompasses linkinggroups which are —CH₂CH₂—, ethylene.

The second category of L units relates to alkyl substituted alkyleneunits chosen from:

-   -   i) —CH(CH₃)CH₂—, 1-methylethylene;    -   ii) —CH₂CH(CH₃)—, 2-methylethylene;    -   iii) —CH(CH₃)CH₂CH₂—, 1-methylpropylene;    -   iv) —CH₂CH(CH₃)CH₂—, 2-methylpropylene; and    -   v) —CH₂C(CH₃)₂CH₂—, 2,2-dimethylpropylene.    -   The first aspect of the second category of L units encompasses        linking groups which are —CH(CH₃)CH₂CH₂—, 1-methylpropylene; as        exemplified in the generic formula:

wherein the above formula encompasses both the R and the S enantiomersof the linking unit.

Synthesis Procedure

The compounds of the present invention can be prepared by the followinggeneral procedure, the formulator adjusting the reaction conditions asis necessary and which one skilled in the art will be able to accomplishwithout undo experimentation.

Step 1: Preparation of intermediate 2-(methylthio)pyrimidine-4(3H)-one

This compound can be used in the preparation of each analog encompassedby the present invention. The general procedure follows.

To a solution of sodium hydroxide (8 g, 200 mmol) in H₂O (75 mL) at roomtemperature is added thiouridine (14.2 g, 100 mmol). The resultingmixture is stirred at room temperature for 20 minutes. Methyl iodide(6.86 mL, 110 mmol) is slowly added dropwise in THF (10 mL) and themixture is stirred at room temperature for 18 hours. A white solid formsupon acidifying the mixture to pH 5 with glacial acetic acid. At thispoint the mixture is cooled in an ice bath and allowed to stand forapproximately 2 hours after which the final product separates as a whitesolid and can be collected by filtration. First crop of crystalstypically yields the desired product in an excess of 60% yield. ¹H NMR(DMSO-d₆, 300 MHz): δ 2.45 (s, 3H), 6.07 (d, J=6.6 Hz, 1H), 7.85 (d,J=6.6 Hz, 1H).

Step 2: Formation of 2-anilino Intermediate

2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one: To a2-(methyl-thio)pyrimidine-4(3H)-one (14.2 g, 100 mmol) in diglyme (60mL) is added the substituted or unsubstituted aniline of choice (200mmol). The resulting mixture is heated to reflux and stirred forapproximately 18 hours. The product which typically forms as a solidupon cooling the mixture to room temperature, is washed with solvent(pentane, hexane, or isopentane). However, solvent can be added to thereaction mixture to induce crystallization if necessary.

Step 3. Formation of 4-chloro-2-anilino Intermediate

2-(Substituted or unsubstituted phenylamino)-4-chloro-pyrimidine: To a2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one (5.02 g,22.6 mmol) and N,N-dimethyl-aniline (450 mL) is added of phosphorusoxychloride (450 mL). The resulting mixture is heated to reflux for 15minutes, cooled to room temperature and concentrated in vacuo. Theresidue is neutralized to pH 7 with 1M NaOH (aqueous). The organic layeris extracted with EtOAc (3×250 mL). The combined organic layers aredried (MgSO₄) and concentrated in vacuo. The residue can be convenientlypurified over silica (5% EtOAc in hexanes) to afford the desiredcompound.

Alternatively, the 4-chloro-2-anilino intermediate can be synthesizedthe following way:

2-(Substituted or unsubstituted phenylamino)-4-chloro-pyrimidine: To a2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one (3.00 g,13.5 mmol) in toluene (30 mL) is added N,N-dimethyl-aniline (3.57 mL,28.4 mmol) and phosphorus oxychloride (1.24 mL, 13.5 mmol). Theresulting mixture is heated to reflux for 15 minutes, cooled to roomtemperature and neutralized to pH 7 with 1M NaOH (aqueous). The organiclayer is extracted with EtOAc (3×250 mL). The combined organic layersare dried (MgSO₄) and concentrated in vacuo. The residue can beconveniently purified over silica (5% EtOAc in hexanes) to afford thedesired compound.

Step 4. Formation of Final Compounds (Analogs) of the Present Invention

To the 2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-oneformed in Step (2) (100 mmol) in THF (500 mL) is addeddiisopropylethylamine (200 mmol) followed by the desired diamine (200mmol). The resulting mixture is heated to reflux for approximately 18hours. The reaction is cooled to room temperature and concentrated invacuo. The residue which forms is diluted with water and extracted withsolvent. The combined organic layers are dried (MgSO₄) and concentratedin vacuo. This residue can be crystallized or purified over silica toafford the final compound.

Schemes I-IV herein below provide illustrative examples of thepreparation of compounds encompassed by the various categories of thepresent invention.

The analogs (compounds) of the present invention are arranged intoseveral categories to assist the formulator in applying a rationalsynthetic strategy for the preparation of analogs which are notexpressly exampled herein. The arrangement into categories does notimply increased or decreased efficacy for any of the compositions ofmatter described herein.

Analog Categories

The compounds which comprise Category I of the present invention are2-(substituted phenylamino)-4-(amino- or substitutedamino-propylene)aminopyrimidines having the formula:

wherein R, R¹, and R² are defined herein below in Table I.

TABLE I No R R¹ R² 1 3-chlorophenyl —H —H 2 3-chlorophenyl —CH₃ —H 33-chlorophenyl —CH₃ —CH₃ 4 3-chlorophenyl —CH₂CH₃ —H 5 3-chlorophenyl—CH₂CH₃ —CH₃ 6 3-chlorophenyl —CH₂CH₃ —CH₂CH₃ 7 4-chlorophenyl —H —H 84-chlorophenyl —CH₃ —H 9 4-chlorophenyl —CH₃ —CH₃ 10 4-chlorophenyl—CH₂CH₃ —H 11 4-chlorophenyl —CH₂CH₃ —CH₃ 12 4-chlorophenyl —CH₂CH₃—CH₂CH₃ 13 3,4-dichlorophenyl —H —H 14 3,4-dichlorophenyl —CH₃ —H 153,4-dichlorophenyl —CH₃ —CH₃ 16 3,4-dichlorophenyl —CH₂CH₃ —H 173,4-dichlorophenyl —CH₂CH₃ —CH₃ 18 3,4-dichlorophenyl —CH₂CH₃ —CH₂CH₃ 193-chloro-4-methylphenyl —H —H 20 3-chloro-4-methylphenyl —CH₃ —H 213-chloro-4-methylphenyl —CH₃ —CH₃ 22 3-chloro-4-methylphenyl —CH₂CH₃ —H23 3-chloro-4-methylphenyl —CH₂CH₃ —CH₃ 24 3-chloro-4-methylphenyl—CH₂CH₃ —CH₂CH₃ 25 3-chloro-2-fluorophenyl —CH₃ —CH₃ 263-chloro-2-fluorophenyl —CH₂CH₃ —CH₃ 27 3-chloro-2-fluorophenyl —CH₂CH₃—CH₂CH₃ 28 3-chloro-4-fluorophenyl —CH₃ —CH₃ 29 3-chloro-4-fluorophenyl—CH₂CH₃ —CH₃ 30 3-chloro-4-fluorophenyl —CH₂CH₃ —CH₂CH₃ 313,4-difluorophenyl —H —H 32 3,4-difluorophenyl —CH₃ —H 333,4-difluorophenyl —CH₃ —CH₃ 34 3,4-difluorophenyl —CH₂CH₃ —H 353,4-difluorophenyl —CH₂CH₃ —CH₃ 36 3,4-difluorophenyl —CH₂CH₃ —CH₂CH₃ 373-trifluoromethylphenyl —H —H 38 3-trifluoromethylphenyl —CH₃ —H 393-trifluoromethylphenyl —CH₃ —CH₃ 40 3-trifluoromethylphenyl —CH₂CH₃ —H41 3-trifluoromethylphenyl —CH₂CH₃ —CH₃ 42 3-trifluoromethylphenyl—CH₂CH₃ —CH₂CH₃ 43 3-trifluoromethyl-4-chlorophenyl —H —H 443-trifluoromethyl-4-chlorophenyl —CH₃ —H 453-trifluoromethyl-4-chlorophenyl —CH₃ —CH₃ 463-trifluoromethyl-4-chlorophenyl —CH₂CH₃ —H 473-trifluoromethyl-4-chlorophenyl —CH₂CH₃ —CH₃ 483-trifluoromethyl-4-chlorophenyl —CH₂CH₃ —CH₂CH₃ 49 3-methoxyphenyl —H—H 50 3-methoxyphenyl —CH₃ —H 51 3-methoxyphenyl —CH₃ —CH₃ 523-methoxyphenyl —CH₂CH₃ —H 53 3-methoxyphenyl —CH₂CH₃ —CH₃ 543-methoxyphenyl —CH₂CH₃ —CH₂CH₃ 55 3-methylphenyl —CH₃ —CH₃ 563-methylphenyl —CH₂CH₃ —CH₃ 57 3-methylphenyl —CH₂CH₃ —CH₂CH₃ 583-ethylphenyl —CH₃ —CH₃ 59 3-ethylphenyl —CH₂CH₃ —CH₃ 60 3-ethylphenyl—CH₂CH₃ —CH₂CH₃

The compounds which comprise Category I of the present invention can beprepared by the procedure outlined herein below in Scheme 1.

EXAMPLE 1N²-(3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine(4)

Preparation of 2-(methylthio)pyrimidine-4(3H)-one (1): To a solution ofsodium hydroxide (6.24 g, 156.07 mmol) in H₂O (55 mL) at roomtemperature is added thiouridine (10 g, 78.03 mmol). The resultingmixture is stirred at room temperature for 20 min. Methyl iodide (5.45mL, 87.40 mmol) in THF (10 mL) is added dropwise slowly and the mixtureis stirred at room temperature for 18 hours. A white solid forms uponacidifying the mixture to pH 5 with glacial acetic acid. The mixture isallowed to stand at 0° C. (ice bath) for 2 hours and filtered to afford7.4 g (67% yield) of the desired compound as a white solid. ¹H NMR(DMSO-d₆, 300 MHz): δ 2.45 (s, 3H), 6.07 (d, J=6.6 Hz, 1H), 7.85 (d,J=6.6 Hz, 1H).

Preparation of 2-(3-chlorophenylamino)pyrimidin-4(3H)-one (2): To2-(methyl-thio)pyrimidine-4(3H)-one, 1, (4.88 g, 34.37 mmol) in diglyme(20 mL) is added 3-chloroaniline (4.3 mL, 68.74 mmol). The resultingmixture is heated to reflux and stirred for 18 hours. A solid forms uponcooling the mixture to room temperature. The solid is washed withhexanes to afford 5.0 g (66% yield) of the desired compound. ¹H NMR(DMSO-d₆, 300 MHz): δ 5.91 (d, J=5.7 Hz, 2H), 7.05 (d, J=7.5 Hz, 1H),7.11 (br s, 1H), 7.32 (t, J=7.8, 15.9 Hz, 1H), 7.45 (d, J=7.8 Hz, 1H),7.86 (d, J=4.5 Hz, 1H), 7.94 (s, 1H).

Preparation of 4-chloro-N-(3-chlorophenyl)pyrimidin-2-amine (3): To2-(3-chlorophenylamino)pyrimidin-4(3H)-one, 2, (3.00 g, 13.5 mmol) intoluene (30 mL) is added N,N-dimethyl-aniline (3.57 mL, 28.4 mmol) andphosphorus oxychloride (1.24 mL, 13.5 mmol). The resulting mixture isheated to reflux for 15 minutes, cooled to room temperature andneutralized to pH 7 with 1M NaOH (aqueous). The organic layer isextracted with EtOAc (3×250 mL). The combined organic layers are dried(MgSO₄) and concentrated in vacuo. The residue is purified over silica(5% EtOAc in hexanes) to afford 2.0 g (61% yield) of the desiredcompound. ¹H NMR (DMSO-d₆, 300 MHz): δ 7.06-7.04 (m, 2H), 7.34 (t,J=8.1, 1H), 7.65-7.61 (m, 1H), 7.93 (m, 1H), 8.50 (d, J=5.1 Hz, 1H),10.26 (s, 1H).

Preparation ofN²-(3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine(4): To 4-chloro-N-(3-chlorophenyl)pyrimidin-2-amine, 3, (0.517 g, 2.16mmol) in THF (10 mL) is added diisopropylethylamine (0.754 mL, 4.32mmol) followed by 3-dimethyl-aminopropylamine (0.544 mL, 4.32 mmol). Theresulting mixture is heated to reflux for 18 hours. The reaction iscooled to room temperature and concentrated in vacuo. The residue isdiluted with 10 mL water and extracted with EtOAc (3×50 mL). Thecombined organic layers are dried (MgSO₄) and concentrated in vacuo.This residue is purified over silica (5% MeOH in CH₂Cl₂ with 0.7% Et₃N)to afford the desired compound as a yellow solid. The compound can beconveniently recrystallized from CH₂Cl₂ and MeOH to afford 0.206 g (40%yield) of a white solid. ¹H NMR (CD₃OD, 300 MHz): δ 1.85-1.95 (m, 2H).2.40 (s, 6H), 2.59-2.65 (m, 2H), 3.40-3.51 (m, 2H), 5.99 (d, J=6.3 Hz,1H), 6.95 (d, J=8.1 Hz, 1H), 7.24 (t, J=8.1 Hz, 1H), 7.40 (d, J=6.9 Hz,1H), 7.78 (d, J=5.7 Hz, 1H), 8.03 (s, 1H). MS (ESI, pos. ion) m/z: 306(M+1).

The following are non-limiting examples of compounds which compriseCategory I of the present invention, the characterization of which willassist the formulator in establishing the chemical formulae of compoundswhich are not specifically exemplified herein.

N²-(3-chlorophenyl)-N⁴-(3-(diethylamino)propyl)pyrimidine-2,4-diamine:¹H NMR (CD₃OD, 300 MHz): δ 1.30 (t, J=7.5 Hz, 6H), 2.15 (m, 2H), 3.18(m, 6H), 3.60 (t, J=5.7 Hz, 1H), 6.15 (d, J=5.7 Hz, 1H), 6.98 (d, J=8.1Hz, 1H), 7.23 (dd, J=8.1, 8.4 Hz, 1H), 7.40 (d, J=8.4 Hz, 1H), 7.81 (brd, J=5.7, 1H), 8.15 (s, 1H). MS (ESI, pos. ion) m/z: 334 (M+1).

N⁴-(3-(dimethylamino)propyl)-N²-(3-methoxyphenyl)pyrimidine-2,4-diaminehydrochloride: ¹H NMR (CD₃OD, 300 MHz): δ 1.27 (t, J=7.5 Hz, 6H),2.03-2.10 (m, 2H), 2.89 (s, 3H), 3.18-3.24 (m, 2H), 3.59-3.63 (m, 2H),3.86 (s, 3H), 6.28 (d, J=7.5 Hz, 1H), 6.90 (d, J=8.4 Hz, 1H), 7.05 (d,J=8.1 Hz, 2H), 7.18 (s, 1H), 7.39 (t, J=7.8 Hz, 1H), 7.67 (d, J=7.2 Hz,1H). MS (ESI, pos. ion) m/z: 302 (M+1).

N⁴-(3-(dimethylamino)propyl)-N²-(3-methylphenyl)pyrimidine-2,4-diaminehydrochloride: ¹H NMR (DMSO-d₆, 300 MHz): δ 1.98 (m, 2H), 2.33 (s, 3H),2.72 (s, 6H), 3.09 (m, 2H), 3.45 (m, 2H), 6.24 (d, J=7.5 Hz, 1H), 6.97(d, J=7.8 Hz, 1H), 7.29 (t, J=7.5 Hz, 1H), 7.40-7.43, (m, 4H), 7.84 (d,J=6.9 Hz, 1H), 9.17 (br sm 1H), 10.47 (br s, 2H). MS (ESI, pos. ion)m/z: 286 (M+1).

N⁴-(3-(dimethylamino)propyl)-N²-(3-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine:¹H NMR (DMSO-d₆, 300 MHz): δ 1.63-1.72 (m, 2H), 2.13 (s, 6H), 2.26-2.31(m, 2H), 3.32-3.34 (m, 2H), 5.98 (d, J=5.7 Hz, 1H), 7.17 (d, J=7.8 Hz,1H), 7.30 (br s, 1H), 7.43 (t, J=7.8 Hz, 1H), 7.83 (br s, 1H), 8.48 (brs, 1H), 9.35 (br s, 1H). ¹⁹F 101.76 (s, 3F). MS (ESI, pos. ion) m/z: 340(M+1).

N²-(3-chlorophenyl)-N⁴-(3-(methylamino)propyl)pyrimidine-2,4-diamine: ¹HNMR (CD₃OD, 300 MHz): δ 2.02-2.11 (m, 2H), 2.72 (s, 3H), 3.06-3.11 (m,2H), 3.60-3.65 (m, 2H), 6.32-6.34 (m, 1H), 7.29-7.51 (m, 3H), 7.72-7.79(m, 2H). MS (ESI, pos. ion) m/z: 292 (M+1).

N⁴-(3-aminopropyl)-N²-(3-chlorophenyl)pyrimidine-2,4-diamine: ¹H NMR(DMSO-d₆, 300 MHz): δ 1.90-1.97 (m, 2H), 2.89 (m, 2H), 3.46-3.52 (m,2H), 6.35 (d, J=7.5 Hz, 1H), 7.22 (d, J=7.8 Hz, 1H), 7.42-7.53 (m, 2H),7.88-7.92 (m, 2H), 8.12 (br s, 3H), 9.62 (br s, 1H), 11.15 (br s, 1H).MS (ESI, pos. ion) m/z: 278 (M+1).

N²-(3,4-difluorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine:¹H NMR (DMSO-d₆, 300 MHz): δ 1.68 (m, 2H), 2.13 (s, 6H), 2.27 (m, 2H),3.32 (m, 2H), 5.95 (d, J=5.4 Hz, 1H), 7.20-7.29 (m, 2H), 7.37-7.41 (m,2H), 7.79 (m, 1H), 8.07-8.14 (m, 1H), 9.18 (s, 1H). ¹⁹F (DMSO-d₆, 300MHz): δ 13.61 (m, 1F), 24.856 (m, 1F). MS (ESI, pos. ion) m/z: 308.27(M+1).

N²-(3-chloro-4-methylphenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine:¹H NMR (CD₃OD, 300 MHz): δ 1.85 (m, 2H), 2.28 (s, 6H), 2.32 (s, 3H),2.48 (t, 2H), 3.46 (m, 2H), 5.96 (d, J=6.3 Hz, 1H), 7.16 (d, J=8.1 Hz,1H), 7.29 (d, J=5.4 Hz, 1H), 7.75 (d, J=5.1 Hz, 1H), 8.00 (s, 1H). MS(ESI, pos. ion) m/z: 320 (M+1).

N²-(3,4-dichlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine:¹H NMR (CDCl₃, 300 MHz): δ 1.85 (m, 2H), 2.34 (s, 6H), 2.52 (t, J=6.0Hz, 2H), 3.50 (m, 2H), 5.91 (d, J=6.0 Hz, 1H), 6.21 (s, 1H), 7.10 (s,1H), 7.30 (d, J=6.0 Hz, 1H), 7.92 (d, J=5.7 Hz, 1H), 8.13 (s, 1H). MS(ESI, pos. ion) m/z: 340 (M+1).

N²-(3-chloro-4-fluorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine:¹H NMR (CDCl₃, 300 MHz): δ 1.79-1.86 (m, 2H), 2.30 (s, 6H), 2.46 (t,J=6.0 Hz, 2H), 3.46 (m, J=6.0 Hz, 2H), 5.89 (d, J=6.0 Hz, 1H), 6.13 (s,1H), 7.06 (t, J=6.0 Hz, 1H), 7.17 (s, 1H), 7.27 (m, 1H), 7.26 (d, J=6.6Hz, 1H), 7.29 (s, 1H), 7.92 (d, J=6.0 Hz, 1H), 8.02 (d, J=6.6 Hz, 1H).HRMS calcd for C₁₅H₁₉ClFN₅ 324.1391 m/z (M+1)⁺; observed 324.1399 m/z.

N²-(4-chloro-3-(trifluoromethyl)phenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine:¹H NMR (CDCl₃, 300 MHz): δ 1.80 (m, 2H), 2.27 (s, 6H), 2.45 (t, J=6.0Hz, 2H), 3.49 (bs, 2H), 5.91 (d, J=6.0 Hz, 1H), 6.37 (t, J=4.8 Hz, 1H)),7.37 (d, J=8.7 Hz, 1H), 7.58 (d, J=8.7 Hz, 1H), 7.94 (d, J=6.0 Hz, 1H),1H), 8.34 (bs, 2H). HRMS calcd for C₁₆H₁₉ClF₃N₅ 374.1359 m/z (M+H)⁺;observed 374.1357 m/z.

N⁴-(3-(dimethylamino)propyl)-N²-(3-ethylphenyl)pyrimidine-2,4-diamine:¹H NMR (CD₃OD, 300 MHz): δ 1.29 (t, J=7.8 Hz, 3H), 2.07 (m, 2H), 2.71(q, J=7.8 Hz, 2H)), 2.85 (s, 6H), 3.16 (m, 2H), 3.60 (t, J=6.6 Hz, 2H),6.24 (d, J=7.5 Hz, 1H), 7.16 (m, 1H), 7.34 (s, 1H), 7.38 (m, 2H), 7.65(d, J=7.2 Hz, 1H). HRMS calcd for C₁₇H₂₅N₅ 300.2188 m/z (M+1)⁺; observed300.2194 m/z.

N²-(2-fluoro-3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine¹H NMR (CDCl₃, 300 MHz): δ 1.75-1.82 (m, 2H), 2.28 (s, 6H), 2.44 (t,J=6.3 Hz, 2H), 3.46 (s, 2H), 5.92 (d, J=6.0 Hz, 1H), 6.02 (bs, 1H),6.97-7.08, (m, 3H), 7.95 (d, J=5.1 Hz, 1H), 8.49 (t, J=7.8 Hz, 1H). HRMScalcd for C₁₅H₁₉N₅FCl, 324.1391 m/z (M+H)⁺; observed 324.1394 m/z.

Further compounds which are encompassed within Category I of the presentinvention but which are not fully exemplified include:

-   -   N⁴-(3-(dimethylamino)propyl)-N²-(2-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(3-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,3-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,4-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,5-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,6-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2-chlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,3-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,4-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,5-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,6-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(4-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,3-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,4-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,5-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(3-(dimethylamino)propyl)-N²-(2,6-dimethylphenyl)pyrimidine-2,4-diamine;        and    -   N⁴-(3-(dimethylamino)propyl)-N²-(3,4-dimethylphenyl)pyrimidine-2,4-diamine.

The compounds which comprise Category II of the present invention are2-(substituted phenylamino)-4-(amino- or substituted amino-ethylene, orsubstituted amino-butylene)-aminopyrimidines having the formula:

wherein R², R³, R⁵, and L are defined herein below in Table II.

TABLE II No. L R² R³ R⁵ 61 —CH₂— —H —H 3-chloro 62 —CH₂— —CH₃ —H3-chloro 63 —CH₂— —CH₃ —CH₃ 3-chloro 64 —CH₂— —CH₂CH₃ —H 3-chloro 65—CH₂— —CH₂CH₃ —CH₃ 3-chloro 66 —CH₂— —CH₂CH₃ —CH₂CH₃ 3-chloro 67—CH₂CH₂— —H —H 3-chloro 68 —CH₂CH₂— —CH₃ —H 3-chloro 69 —CH₂CH₂— —CH₃—CH₃ 3-chloro 70 —CH₂CH₂— —CH₂CH₃ —H 3-chloro 71 —CH₂CH₂— —CH₂CH₃ —CH₃3-chloro 72 —CH₂CH₂— —CH₂CH₃ —CH₂CH₃ 3-chloro 73 —CH₂C(CH₃)₂CH₂— —H —H3-trifluoromethyl 74 —CH₂C(CH₃) ₂CH₂ — —CH₃ —H 3-trifluoromethyl 75—CH₂C(CH₃) ₂CH₂ — —CH₃ —CH₃ 3-trifluoromethyl 76 —CH₂C(CH₃) ₂CH₂ ——CH₂CH₃ —H 3-trifluoromethyl 77 —CH₂C(CH₃) ₂CH₂ — —CH₂CH₃ —CH₃3-trifluoromethyl 78 —CH₂C(CH₃)₂CH₃₂— —CH₂CH₃ —CH₂CH₃ 3-trifluoromethyl79 —CH₂CH₂CH₂CH₂— —H —H 3-chloro 80 —CH₂CH₂CH₂CH₂— —CH₃ —H 3-chloro 81—CH₂CH₂CH₂CH₂— —CH₃ —CH₃ 3-chloro 82 —CH₂CH₂CH₂CH₂— —CH₂CH₃ —H 3-chloro83 —CH₂CH₂CH₂CH₂— —CH₂CH₃ —CH₃ 3-chloro 84 —CH₂CH₂CH₂CH₂— —CH₂CH₃—CH₂CH₃ 3-chloro

The compounds which comprise Category II of the present invention can beprepared by the procedure outlined herein below in Schemes II and IIIand Examples 3 and 4.

EXAMPLE 2N⁴-[3-(dimethylamino)-2,2-dimethylpropyl]-N²-[3-(trifluoromethyl)phenyl]pyrimidine-2,4-diamineHydrochloride (7)

Preparation of 2-(3-trifluoromethylphenylamino)pyrimidin-4(3H)-one (5):To 2-(methyl-thio)pyrimidine-4(3H)-one, 1, (5 g, 35.21 mmol) in diglyme(25 mL) is added 3-trifluoromethylaniline (5.26 mL, 42.25 mmol). Theresulting mixture is heated to reflux and stirred for 18 hours. A solidforms upon cooling the mixture to room temperature. The solid is washedwith hexanes to afford 1.9 g (21% yield) of the desired compound. MS(ESI, pos. ion) m/z: 256 (M+1).

Preparation of 4-chloro-N-(3-trifluoromethylphenyl)pyrimidin-2-amine(6): To 2-(3-chloro-phenylamino)pyrimidin-4(3H)-one, 5, (1.9 g, 7.4mmol) and N,N-dimethyl-aniline (2 mL) is added of phosphorus oxychloride(20 mL). The resulting mixture is heated to reflux for 15 minutes,cooled to room temperature and concentrated in vacuo. The residue isneutralized to pH 7 with 1M NaOH (aqueous). The organic layer isextracted with ethyl acetate (3×50 mL). The combined organic layers aredried (MgSO₄) and concentrated in vacuo. The residue is purified oversilica (5% EtOAc in hexanes) to afford 0.62 g (31% yield) of the desiredcompound. ¹H NMR (DMSO-d₆, 300 MHz): δ 7.07 (d, J=6.3 Hz, 1H), 7.34 (d,J=8.4 Hz, 1H), 7.55 (t, J=8.1 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 8.20 (s,1H), 8.52 (d, J=6.0 Hz, 1H), 10.40 (s, 1H). ¹⁹F NMR (DMSO-d₆, 282 MHz):δ 101.67 (s, 3F).

Preparation ofN⁴-[3-(dimethylamino)-2,2-dimethylpropyl]-N²-[3-(trifluoro-methyl)phenyl]pyrimidine-2,4-diaminehydrochloride (7):4-Chloro-N-(3-(trifluoro-methyl)phenyl)pyrimidin-2-amine, 6, (200 mg,0.73 mmol), N¹,N¹,2,2-tetramethyl-propane-1,3-diamine (0.232 mL, 1.46mmol), and diisopropylethylamine (0.254 mL, 1.46 mmol) are dissolved inTHF (5 mL) and heated to reflux for 20 hours. The reaction is cooled toroom temperature and partitioned between EtOAc and water. The organiclayer is dried (MgSO₄), concentrated in vacuo, and purified over silica(5% MeOH ramped to 6% MeOH in CH₂Cl₂ with 0.7% added triethylamine) togive an oil. The oil is dissolved in MeOH (0.5 mL) and Et₂O (20 mL) andcooled to 0° C. To this solution is added a solution of 4M HCl indioxane (0.25 mL) in one portion. The white precipitate which forms iscollected by filtration and dried at reduced pressure to afford thedesired compound as a white solid: ¹H NMR (DMSO-d₆, 300 MHz) δ 1.09 (s,6H), 2.81 (s, 3H), 2.83 (s, 3H), 3.11 (d, J=4.2 Hz, 2H), 3.38-3.50 (m,2H), 6.48 (d, J=7.2 Hz, 1H), 7.51 (d, J=7.5 Hz, 1H), 7.66 (t, J=7.5 Hz,1H), 7.74 (d. J=6.6 Hz, 1H), 7.91 (d, J=7.2 Hz, 1H), 8.20 (s, 1H), 9.67(bs, 1H), 10.80 (bs, 1H); HRMS calcd for C₁₈H₂₄F₃N₅, 368.2062 m/z(M+H)⁺; observed 368.2072 m/z.

EXAMPLE 3N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine(8)

Preparation ofN²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine(8):To 4-chloro-N-(3-chlorophenyl)pyrimidin-2-amine (0.2 g, 0.84 mmol) inTHF (4 mL) is added diisopropylethylamine (0.29 mL, 1.67 mmol) followedby 4-dimethylaminobutylamine (0.2 g, 1.67 mmol). The resulting mixtureis heated to reflux for 6 hours. Another 2 equivalents of4-dimethylaminobutylamine (0.2 g, 1.67 mmol) are added and the reactionheated at reflux for 18 hours. The reaction is cooled to roomtemperature and concentrated in vacuo. The residue is diluted with water(5 mL) and extracted with EtOAc (3×25 mL). The combined organic layersare dried (MgSO₄) and concentrated in vacuo. This residue is purifiedover silica (5% MeOH in CH₂Cl₂ with 0.7% Et₃N) to afford 7 mg (3% yield)of the desired compound. ¹H NMR (CD₃OD, 300 MHz): δ 1.62-1.66 (m, 4H),2.26 (s, 6H), 2.39-2.44 (m, 2H), 3.46-3.48 (m, 2H), 5.97 (d, J=5.7 Hz,1H), 6.93 (d, J=8.1 Hz, 1H), 7.22 (t, J=8.1 Hz, 1H), 7.40 (d, J=8.1 Hz,1H), 7.75 (d, J=5.7 Hz, 1H), 8.06 (s, 1H). MS (ESI, pos. ion) m/z: 320(M+1).

The following are non-limiting examples of compounds which compriseCategory II of the present invention, the characterization of which willassist the formulator in establishing the chemical formulae of compoundswhich are not specifically exemplified herein.

N²-(3-chlorophenyl)-N⁴-(2-(dimethylamino)ethyl)pyrimidine-2,4-diamine.¹H NMR (CD₃OD, 300 MHz): δ 2.33 (s, 6H), 2.62 (t, J=6.0 Hz, 2H), 3.58(t, J=6.0 Hz, 2H), 5.99 (d, J=6.0 Hz, 1H), 6.95 (d, J=8.1 Hz, 1H), 7.23(t, J=8.1 Hz, 1H), 7.44 (d, J=8.1 Hz, 1H), 7.78 (d, J=6.0 Hz, 1H), 7.95(s, 1H). MS (ESI, pos. ion) m/z: 292 (M+1).

N⁴-(3-aminopropyl)-N²-(3-chlorophenyl)-N⁴-methylpyrimidine-2,4-diamine:¹H NMR (DMSO-d₆, 300 MHz): δ 1.88-1.92 (m, 2H), 2.78-2.84 (m, 2H), 3.13(s, 3H), 3.62-3.68 (m, 2H), 3.43 (br s, 1H), 7.06-7.08 (m, 1H),7.30-7.38 (m, 1H), 7.50-7.53 (m, 1H), 7.93-8.04 (m, 4H), 10.25 (br s,1H). MS (ESI, pos. ion) m/z: 292 (M+1).

Further compounds which are encompassed within Category II of thepresent invention but which are not fully exemplified include:

-   -   N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(3-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,3-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,4-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,5-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,6-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2-chlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,3-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,4-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,5-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,6-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(3-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(4-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,3-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,4-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,5-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,6-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(2-(dimethylamino)ethyl)-N²-(3,4-dimethylphenyl)pyrimidine-2,4-diamine;    -   N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(3-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,3-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,4-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,5-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,6-difluorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2-chlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(4-chlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,3-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,4-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,5-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,6-dichlorophenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(4-methylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,3-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,4-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,5-dimethylphenyl)pyrimidine-2,4-diamine;    -   N⁴-(4-(dimethylamino)butyl)-N²-(2,6-dimethylphenyl)pyrimidine-2,4-diamine;        and    -   N⁴-(4-(dimethylamino)butyl)-N²-(3,4-dimethylphenyl)pyrimidine-2,4-diamine.

The compounds which comprise Category III of the present invention are2-(substituted phenylamino)-4-(amino- or substitutedamino-propylene)aminopyrimidines having the formula:

wherein R⁵ is substituted or unsubstituted aryl, substituted orunsubstituted heteroaryl, or substituted or unsubstituted heterocyclic,and R¹, and R² are defined herein below in Table IIII.

TABLE III No R⁵ R¹ R² 85 3-phenyl —H —H 86 3-phenyl —CH₃ —H 87 3-phenyl—CH₃ —CH₃ 88 3-phenyl —CH₂CH₃ —H 89 3-phenyl —CH₂CH₃ —CH₃ 90 3-phenyl—CH₂CH₃ —CH₂CH₃ 91 4-phenyl —H —H 92 4-phenyl —CH₃ —H 93 4-phenyl —CH₃—CH₃ 94 4-phenyl —CH₂CH₃ —H 95 4-phenyl —CH₂CH₃ —CH₃ 96 4-phenyl —CH₂CH₃—CH₂CH₃ 97 thiazol-4-yl —H —H 98 thiazol-4-yl —CH₃ —H 99 thiazol-4-yl—CH₃ —CH₃ 100 thiazol-4-yl —CH₂CH₃ —H 101 thiazol-4-yl —CH₂CH₃ —CH₃ 102thiazol-4-yl —CH₂CH₃ —CH₂CH₃ 103 2-methylthiazol-4-yl —H —H 1042-methylthiazol-4-yl —CH₃ —H 105 2-methylthiazol-4-yl —CH₃ —CH₃ 1062-methylthiazol-4-yl —CH₂CH₃ —H 107 2-methylthiazol-4-yl —CH₂CH₃ —CH₃108 2-methylthiazol-4-yl —CH₂CH₃ —CH₂CH₃ 109 imidazol-2-yl —H —H 110imidazol-2-yl —CH₃ —H 111 imidazol-2-yl —CH₃ —CH₃ 112 imidazol-2-yl—CH₂CH₃ —H 113 imidazol-2-yl —CH₂CH₃ —CH₃ 114 imidazol-2-yl —CH₂CH₃—CH₂CH₃ 115 imidazol-4-yl —H —H 116 imidazol-4-yl —CH₃ —H 117imidazol-4-yl —CH₃ —CH₃ 118 imidazol-4-yl —CH₂CH₃ —H 119 imidazol-4-yl—CH₂CH₃ —CH₃ 120 imidazol-4-yl —CH₂CH₃ —CH₂CH₃ 121 2-methylimidazol-4-yl—H —H 122 2-methylimidazol-4-yl —CH₃ —H 123 2-methylimidazol-4-yl —CH₃—CH₃ 124 2-methylimidazol-4-yl —CH₂CH₃ —H 125 2-methylimidazol-4-yl—CH₂CH₃ —CH₃ 126 2-methylimidazol-4-yl —CH₂CH₃ —CH₂CH₃

The compounds which comprise Category III of the present invention canbe prepared by the following procedures outlined herein below in SchemeIV and V and Examples 4 and 5.

EXAMPLE 4N⁴-(3-(dimethylamino)propyl)-N²-m-biphenylpyrimidine-2,4-diaminehydrochloride (11)

Preparation of 2-(3-biphenylamino)pyrimidin-4(3H)-one (9): To2-(methylthio)-pyrimidine-4(3H)-one, 1, (790 mg, 5.5 mmol) in diglyme (5mL) is added 3-amino-biphenyl (1.91 g, 11.2 mmol). The resulting mixtureis stirred at reflux for 18 hours. The mixture is cooled to roomtemperature and hexane is added and a precipitate forms which iscollected by filtration to afford 1.34 g (92% yield) of the desiredcompound which is used without further purification. MS (ESI, pos. ion)m/z: 264 (M+1).

Preparation of 4-chloro-N-(3-biphenyl)pyrimidin-2-amine (10): To2-(3-biphenyl-amino)pyrimidin-4(3H)-one, 9, (1.34 g, 5.0 mmol), andN,N-dimethylaniline (1.5 mL) is added phosphorus oxychloride (10 mL).The resulting mixture is heated to reflux, stirred for 1 hour, cooled toroom temperature and concentrated in vacuo. The residue is neutralizedto pH 7 with 1M NaOH (aqueous). The organic layer is extracted withEtOAc (2×50 mL). The combined organic layers are washed once with brine,dried (MgSO₄), and concentrated in vacuo. The residue is purified oversilica (5% EtOAc in hexanes) to afford 780 mg (54% yield) of the desiredcompound. MS (ESI, pos. ion) m/z: 282 (M+1).

Preparation ofN²-(3-biphenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diaminehydrochloride (11): To 4-chloro-N-(3-biphenyl)pyrimidin-2-amine, 10,(102 mg, 0.38 mmol) in THF (5 mL) is added N,N-diisopropylethylamine(0.15 mL, 0.84 mmol) followed by 3-(dimethylamino)propylamine (0.15 mL,1.2 mmol). The resulting mixture is heated to reflux for 48 hours. Thereaction is cooled to room temperature and concentrated in vacuo toafford a residue which is purified over silica (95:3:2 CH₂Cl₂/MeOH/7NNH₃ in MeOH) to afford 61 mg (48% yield) of the desired compound. Thehydrochloride salt can be formed by treatment of the neutral compoundwith HCl in dioxane (4M). ¹H NMR (CD₃OD, 300 MHz): 6(ppm) 1.74 (m, 2H),2.15 (s, 6H), 2.27 (t, J=6.0 Hz, 2H), 3.40 (m, 2H), 5.94 (d, J=6.0 Hz,1H), 7.19 (d, J=7.2 Hz, 1H), 7.34 (m, 2H), 7.42 (m, 2H), 7.53 (d, J=7.5Hz, 1H), 7.61 (m, 2H), 7.76 (d, J=5.4 Hz, 1H), 8.12 (s, 1H). MS (ESI,pos. ion) m/z: 348 (M+1).

EXAMPLE 5N⁴-(3-(dimethylamino)propyl)-N²-(3-(2-methylthiazol-4-yl)phenyl)pyrimidine-2,4-diamineHydrochloride (13)

Preparation of2-(3-(2-methylthiazol-4-yl)phenylamino)pyrimidin-4(3H)-one (11): To2-(methyl-thio)pyrimidine-4(3H)-one, 1, (1 g, 14.08 mmol) in diglyme (9mL) is added 3-(2-methylthiazol-4-yl)benzenamine (3.20 g, 16.90 mmol).The resulting mixture is heated to reflux and stirred for 18 hours. Asolid forms upon cooling the mixture to room temperature. The solid iswashed with hexanes to afford 1.0 g (25% yield) of the desired compound.MS (ESI, pos. ion) m/z: 285 (M+1).

Preparation of4-chloro-N-(3-(2-methylthiazol-4-yl)phenyl)pyrimidin-2-amine (12): To2-(3-(2-methylthiazol-4-yl)phenylamino)pyrimidin-4(3H)-one, 11, (1.0 g,3.5 mmol) and N,N-dimethyl-aniline (1 mL) is added phosphorusoxychloride (9.5 mL). The resulting mixture is heated to reflux for 15minutes, cooled to room temperature and concentrated in vacuo. Theresidue is neutralized to pH 7 with 1M NaOH (aqueous). The organic layeris extracted with ethyl acetate (3×50 mL). The combined organic layersare dried (MgSO₄) and concentrated in vacuo. The residue is purifiedover silica (5% EtOAc in hexanes) to afford 0.50 g (47% yield) of thedesired compound. ¹H NMR (DMSO-d₆, 300 MHz): δ 2.73 (s, 3H), 6.98 (d,J=5.1 Hz, 1H), 7.37 (t, J=8.1 Hz, 1H), 7.57 (d, J=8.1 Hz, 1H), 7.72 (d,J=8.1 Hz, 1H), 7.83 (s, 1H), 8.25 (s, 1H), 8.46 (d, J=5.1 Hz, 1H), 10.08(s, 1H). MS (ESI, pos. ion) m/z: 303 (M+1).

Preparation ofN⁴-(3-(dimethylamino)propyl)-N²-(3-(2-methylthiazol-4-yl)phenyl)pyrimidine-2,4-diaminehydrochloride (13):4-chloro-N-(3-(2-methylthiazol-4-yl)phenyl)pyrimidin-2-amine, 12, (400mg, 1.30 mmol), N¹,N¹-dimethylpropane-1,3-diamine (0.25 mL, 2.0 mmol),and diisopropylethylamine (0.46 mL, 2.6 mmol) are dissolved in THF (10mL) and heated to reflux for 20 hours. The reaction is cooled to roomtemperature and partitioned between EtOAc and water. The organic layeris dried (MgSO₄), concentrated in vacuo, and purified over silica (5%MeOH ramped to 6% MeOH in CH₂Cl₂ with 0.7% added triethylamine) to givean oil. The oil is dissolved in MeOH (0.5 mL) and Et₂O (20 mL) andcooled to 0° C. To this solution is added a solution of 4M HCl indioxane (0.25 mL) in one portion. The white precipitate which forms iscollected by filtration and dried at reduced pressure to afford thedesired compound as a white solid: ¹H NMR (CD₃OD, 300 MHz) δ 2.03-2.08(m, 2H), 2.79 (s, 6H), 2.80 (s, 3H), 3.10-3.15 (m, 2H), 3.65 (t, J=6.6Hz, 2H), 6.30 (d, J=7.5 Hz, 1H), 7.45 (d, J=7.8 Hz, 1H), 7.54 (t, J=7.8Hz, 1H), 7.70 (d. J=7.5 Hz, 1H), 7.77 (s, 1H), 7.82 (d, J=7.8 Hz, 1H),8.19 (s, 1H); HRMS calcd for C₁₉H₂₄N₆S, 369.1861 m/z (M+H)⁺; observed369.1855 m/z.

The compounds of the present invention are inhibitors of Protein KinaseC-alpha (PKC-α), therefore, they are PKC-α inhibitors which are capableof improving myocardial contraction and relaxation performance and slowthe progression of heart failure. The compounds potentially inhibitadditional isoforms of conventional PKC, such as PKC-β or PKc-γ. This isnot undesired and can lead to increased pharmacological effects.

The level of disease, for example, the relative degree of heart failuredue to PKC-α activity will vary from patient to patient and bepredicated by other exacerbating circumstances, inter alia, presence ofother disease conditions (diabetes, high blood pressure, and the like)or patients may suffer from other conditions such as obesity. Therefore,the formulator may be required to employ differing levels or amounts ofthe compounds described herein to obtain a therapeutic level. Theformulator can determine this amount by any of the known testingprocedures known to the artisan.

Formulations

The present invention also relates to compositions or formulations whichcomprise the PKC-α inhibitors according to the present invention. Ingeneral, the compositions of the present invention comprise:

-   -   a) an effective amount of one or more        2-arylamino-4-(aminoalkylene)amino-pyrimidines or salts thereof        according to the present invention which are effective for        inhibiting PKC-α; and    -   b) one or more excipients.    -   For the purposes of the present invention the term “excipient”        and “carrier” are used interchangeably throughout the        description of the present invention. One aspect of excipient        and carrier relate to their definition in terms of a medicament,        said terms are defined in that respect as, “ingredients which        are used in the practice of formulating a safe and effective        pharmaceutical composition.”

The formulator will understand that excipients are used primarily toserve in delivering a safe, stable, and functional pharmaceutical,serving not only as part of the overall vehicle for delivery but also asa means for achieving effective absorption by the recipient of theactive ingredient. An excipient may fill a role as simple and direct asbeing an inert filler, or an excipient as used herein may be part of apH stabilizing system or coating to insure delivery of the ingredientssafely to the stomach. The formulator can also take advantage of thefact the compounds of the present invention have improved cellularpotency, pharmacokinetic properties, as well as improved oralbioavailability.

Non-limiting examples of compositions according to the present inventioninclude:

-   -   a) from about 0.001 mg to about 1000 mg of one or more PKC-α        inhibitors according to the present invention; and    -   b) one or more excipient.

Another embodiment according to the present invention relates to thefollowing compositions:

-   -   a) from about 0.01 mg to about 100 mg of one or more PKC-α        inhibitors according to the present invention; and    -   b) one or more excipient.

A further embodiment according to the present invention relates to thefollowing compositions:

-   -   a) from about 0.1 mg to about 10 mg of one or more PKC-α        inhibitors according to the present invention; and    -   b) one or more excipient.

The term “effective amount” as used herein means “an amount of one ormore PKC-α inhibitors, effective at dosages and for periods of timenecessary to achieve the desired result.” An effective amount may varyaccording to factors known in the art, such as the disease state, age,sex, and weight of the human or animal being treated. Althoughparticular dosage regimes may be described in examples herein, a personskilled in the art would appreciated that the dosage regime may bealtered to provide optimum therapeutic response. For example, severaldivided doses may be administered daily or the dose may beproportionally reduced as indicated by the exigencies of the therapeuticsituation. In addition, the compositions of the present invention can beadministered as frequently as necessary to achieve a therapeutic amount.

Method of Use

The present invention also relates to a method for improving cardiaccontraction/relaxation parameters in heart failure patients and/orattenuating adverse cardiac remodeling and prevent or slow theprogression of worsening heart failure. The present method comprises thestep of administering to a human or higher mammal an effective amount ofa composition comprising one or more of the PKC-α inhibitors accordingto the present invention.

A present invention also relates to a method of treating or preventingchronic or acute heart failure, said method comprised of the step ofadministering to a patient in need thereof a pharmaceutically acceptableamount of a compound according to claim 1 or a therapeuticallyacceptable salt thereof in combination with a drug selected that acts onthe renin-angiotensin-aldosterone system, a diuretic, digoxin or aβ-adrenergic receptor blockers, natriuretic peptides and inotropicagents.

The present invention also relates to the use of the2-arylamino-4-(amino-alkylene)amino-pyrimidines or salts thereof,according to the present invention in the manufacture of a medicamentfor the treatment of heart disease wherein inhibition of PKC-α providesa benefit.

Procedures Assessment of PKC-α Inhibitory Activity

Measurement of PKC-α enzyme activity is performed using full-lengthhuman PKC-α enzyme (Upstate Biotechnology) at a final concentration of0.12 μg/ml in a kinase assay buffer (0.09 mg/ml bovine serum albumin(BSA), 210 μM ethylenediaminetetraacetic acid (EDTA), 360 μM CaCl₂, 1 mMTris-HCl, pH=7.5, 0.5 mM MgCl₂, 0.015 mg/ml phosphatidylserine and 0.015mg/ml diacylglycerol). The reaction is initiated by addition ofadenosine triphosphate (ATP; final concentration 45 μM) and a peptidesubstrate consisting of amino acids 28-43(Ala-Ala-Lys-Ile-Gln-Ala-Ser-Phe-Arg-Gly-His-Met-Ala-Arg-Lys-Lys) ofneurogranin (Promega; final concentration 22 μM). After a 30 minuteincubation at 24° C. the reaction is terminated by adding 5 μL of thereaction mixture into 50 μL of MALDI matrix solution (5 mg/mlα-cyano-4-hydroxycinnamic acid in 50% Acetonitrile/H₂O, 0.1% TFA, 5 mMammonium phosphate). Two microliters of the stopped reaction mixture istransferred onto a MALDI-TOF mass spectrometer target plate.

All spectra are collected on an Applied Biosystems 4700 ProteomicsAnalyzer MALDI-TOF MS equipped with a Nd:YAG laser (355 nm, 3 ns pulsewidth, 200 Hz repetition rate) in negative ion reflector mode. Thesystem is operated with 4700 Explorer software, version 3.0. Automatedacquisition parameters are adjusted to capture and average only thoseindividual spectra within defined success criteria. Specifically, signalintensities for the substrate peptide are set to a minimum threshold of3000 counts and a maximum intensity of 65,000 counts. This ensured thatneither null spectra nor saturated spectra are averaged into the finalreadout. Between 1000 and 1500 laser shots are averaged for each sample.Data are collected in triplicate from 3 successive days to capture themaximum variability related to preparation of enzyme reaction, transferof samples to MALDI target plates, data collection, and data extraction.

The isotope cluster areas for each peptide substrate and product peaksare extracted into a Microsoft Excel worksheet from the 10×10 array ofspectral data simultaneously using the automated analysis functionprovided within the 4700 Explore software. The isotope cluster area isdefined by the software algorithm based on the molecular weight and thegeneral elemental composition of the peptides. The percent conversion (%C) of substrate to product is calculated as the cluster area of theproduct (P) divided by the sum of the cluster areas of the substrate (S)and the product multiplied by 100 as represented by the followingequation:

${\%\mspace{14mu} C} = {\frac{P}{P + S} \times 100.}$

For dose-dependent inhibition studies, the inhibition is plotted as a %Maximal Activity (% MA). Equation one is a measure of the ratio ofproduct to substrate then solving for the % C. However, to measureinhibition of the enzyme activity, one must measure the degree to whichthat activity (% C) is curtailed. Thus the dose-dependant inhibitiondata is plotted as % MA where the maximal activity is the % C measuredin control reactions with no inhibitor as represented by the followingequation:

${\%\mspace{14mu}{MA}} = {\frac{{\%\mspace{14mu} C\mspace{14mu}{with}\mspace{14mu}{inhibitor}}\mspace{14mu}}{\%\mspace{14mu} C\mspace{14mu}{with}\mspace{14mu}{no}\mspace{14mu}{inhibitor}} \times 100.}$

Evaluation of PKCα Inhibitors in Cardiomyocytes

Determination of PKCα activity in cells is determined using murine HL-1atrial cardiac muscle cells. On day 1, HL-1 cells are plated at 18,000cells/well in a 96-well tissue culture plate. Cells are cultured in 0.1ml Claycomb growth medium (without norepinephrine) supplemented with 10%fetal bovine serum, 200 mM glutamine and 1% antibiotic/antimycotic. Onday 2, cells are washed 1× with 100 μl of phosphate buffered saline(PBS) and the medium is replaced with 100 μl serum free Claycomb mediumsupplemented with 200 mM glutamine. For compound testing, the medium isremoved and replaced with serum free Claycomb medium supplemented with200 mM glutamine containing different concentrations of compound at afinal volume of 50 μl. Compounds are dissolved in 100% dimethylsulfoxide(DMSO) and final DMSO concentrations are maintained at 0.5%. Plates arethen incubated for 30 minutes at 37° C. in a 5% CO₂ incubator. Themedium is then removed and the plates are rinsed 1× with ice-cold 100 μlPBS. The PBS is removed and replaced with 10 μl of ice-cold lysis bufferconsisting of B-PERII detergent (Pierce) diluted 1:1 in distilled waterand including a final concentration of 0.3% β-mercaptoethanol, 50 μg/mlphenylmethylsulfonylfluoride (PMSF) 10 mM benzamidine, 10 nM okadaicacid, 20 μg/ml leupeptin and 20 μg/ml soybean trypsin inhibitor. Theplates are gently mixed for 10-20 minutes at 4° C. Next, 90 μl ofcoactivation buffer, consisting of 0.1 mg/ml BSA, 250 μM EDTA, 400 μMCaCl₂, is added to each well. Twenty-five microliters of the celllysate/coactivation buffer solution is removed from each well and theenzyme activity measured by addition of 25 μl of substrate solution,consisting of 0.1 mg/ml bovine serum albumin, 235 μM EDTA, 400 μM CaCl₂,1 mM Tris-HCl, pH=7.5, 0.5 mM MgCl₂, 0.015 mg/ml phosphatidylserine and0.015 mg/ml diacylglycerol, 20 μM ATP and 2 μM of an octapeptidefragment of the EGF receptor (Arg-Lys-Arg-Thr-Leu-Arg-Arg-Leu). After a30 minute incubation at 24° C. the reaction is terminated by adding 5 μLof the reaction mixture into 50 μL of MALDI matrix solution (5 mg/mlα-cyano-4-hydroxycinnamic acid in 50% Acetonitrile/H₂O, 0.1% TFA, 5 mMammonium phosphate). Two microliters of the stopped reaction mixture istransferred onto a MALDI-TOF mass spectrometer target plate.Dose-dependent inhibition is measured by mass spectrometry as % maximalactivity as described above for the isolated PKCα inhibition assays.

In Vivo Evaluation of PKCα Inhibitors in the Anesthetized Rat

Selected PKCα inhibitors are evaluated in rats with acute heart failure(HF) after myocardial infarction (MI) for effects on cardiaccontractility and hemodynamics. Male, Sprague-Dawley rats areanesthetized with isoflurane, intubated, placed on ventilators andmaintained at a surgical plane of anesthesia during the course of theexperiment. The animals are instrumented, for the measurement of leftventricular function (+dP/dt, LVDP), arterial blood pressure, and theECG is monitored for the incidence of arrhythmias. A thoracotomy isperformed at the fourth intercostal space to visualize the heart, thepericardium is opened and a suture is placed around the left anteriordescending (LAD) coronary artery approximately 3-4 mm from its origin.When hemodynamic values are stabilized, the LAD is permanently ligatedto induce a myocardial infarction. Severe arrhythmia are treated withthe administration of lidocaine. Typically, cardiac function stabilizedapproximately 40-60 min after ligation and baseline hemodynamic valuesare measured.N²-(3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine,100 and 300 nmol/kg/min for 10 min each dose, and hemodynamic parametersare measured after each dose. The effects of treatment are normalized topre-treatment baseline values and expressed as a percentage. Statisticalsignificance (p<0.05) is evaluated using one-way ANOVA and Dunnett'smultiple comparison test.

In Vivo Evaluation of PKCα Inhibitors in the Anesthetized Rat

Selected PKCα inhibitors are evaluated in rats with myocardialinfarction (MI) for effects on cardiac contractility and hemodynamics.

Male, Sprague-Dawley or Lewis rats weighing between 225-500 gm areanesthetized with isoflurane and an MI is induced as follows. Athoracotomy is performed at the fourth intercostal space to visualizethe heart, the pericardium is opened and a suture is placed around theleft anterior descending (LAD) coronary artery approximately 3-4 mm fromits origin. When hemodynamic values are stabilized, the LAD ispermanently ligated to induce a myocardial infarction. Severearrhythmias are treated with the administration of lidocaine. Typically,cardiac function stabilized approximately 40-60 min after ligation andbaseline hemodynamic values are measured.

The effects of inhibitors on cardiac contractility and hemodynamics areevaluated in MI rats as follows. The animals are anesthetized withisoflurane. A femoral artery is isolated and cannulated for themeasurement of systemic blood pressure. A jugular vein is isolated andcannulated for the intravenous infusion of inhibitor. The right carotidartery is isolated and a Millar conductance catheter is inserted to theleft ventricle (LV) of the heart. The LV systolic pressure,end-diastolic pressure, +dP/dt_(max), −dP/dt_(min), and heart rate arederived from the LV pressure waveform. Mean arterial blood pressure isderived from the systemic blood pressure waveform. Data are recordedcontinuously and derived using computerized data acquisition software(Notocord or Powerlab). After a period of stabilization, PKC-αinhibitors are infused at the following infusion doses in MI rats: 10,30, 100, 300 and 1000 nmol/kg/min. The infusion of each dose is allowedto run for at least five minutes. At the end of the test infusions, 5.0μg/kg/min of dobutamine is infused. The effects of treatment arenormalized to pretreatment baseline values and expressed as apercentage. Statistical significance (p<0.05) is evaluated using aone-way ANOVA and Dunnett's multiple comparison test.

Table IV provides non-limiting examples of PKC-α IC₅₀ values forrepresentative compounds of the present invention.

TABLE IV PKC-α IC₅₀ No. Compound (nM) 1N²-(3-chlorophenyl)-N⁴-(3-aminopropyl)pyrimidine-2,4- 312 diamine 2N²-(3-chlorophenyl)-N⁴-[3- 4 (methylamino)propyl]pyrimidine-2,4-diamine3 N²-(3-chlorophenyl)-N⁴-[3- 0.8(dimethylamino)propyl]pyrimidine-2,4-diamine 6N²-(3-chlorophenyl)-N⁴-[3- 1 (diethylamino)propyl]pyrimidine-2,4-diamine15 N²-(3,4-dichlorophenyl)-N⁴-[3- 15(dimethylamino)propyl]pyrimidine-2,4-diamine 21N²-(3-chloro-4-methylphenyl)-N⁴-[3- 3(dimethylamino)propyl]-pyrimidine-2,4-diamine 25N²-(3-chloro-2-fluorophenyl)-N⁴-[3- 23(dimethylamino)propyl]-pyrimidine-2,4-diamine 27N²-(3-chloro-4-fluorophenyl)-N⁴-[3- 8(dimethylamino)propyl]-pyrimidine-2,4-diamine 33N²-(3,4-difluorophenyl)-N⁴-[3-(dimethylamino)propyl]- 65pyrimidine-2,4-diamine 39 N²-(3-trifluoromethylphenyl)-N⁴-[3- 0.5(dimethylamino)propyl]-pyrimidine-2,4-diamine 45N²-(3-trifluoromethyl-4-chlorophenyl)-N⁴-[3- 3(dimethylamino)-propyl]pyrimidine-2,4-diamine 51N²-(3-methoxyphenyl)-N⁴-[3- 14d(dimethylamino)propyl]pyrimidine-2,4-iamine 55N²-(3-methylphenyl)-N⁴-[3- 40(dimethylamino)propyl]pyrimidine-2,4-diamine 58N²-(3-methylphenyl)-N⁴-[3- 93(dimethylamino)propyl]pyrimidine-2,4-diamine 69N²-(3-chlorophenyl)-N⁴-[2- 83(dimethylamino)ethyl]pyrimidine-2,4-diamine 81N²-(3-chlorophenyl)-N⁴-[4- 12(dimethylamino)butyl]pyrimidine-2,4-diamine 87 N²-(biphenyl-3-yl)-N⁴-[3-0.4 (dimethylamino)propyl]pyrimidine-2,4-diamine 105N²-[3-(2-methyl-thiazol-4-yl)phenyl]-N⁴-[3- 48(dimethylamino)-propyl]pyrimidine-2,4-diamine

While particular embodiments of the present invention have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this invention.

1. A compound having the formula:

wherein R is a unit having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄substituted or unsubstituted linear, branched, or cyclic alkyl; L is alinking unit having the formula:—[C(R^(4a)R^(4b))]_(n) — each R^(4a) and R^(4b) is independently chosenfrom: i) hydrogen; or ii) C₁-C₄ linear, branched, or cyclic alkyl; theindex n is from 1 to 4; and wherein R¹ is chosen from 3-chlorophenyl,4-chlorophenyl, 3,4-dichlorophenyl, 3-chloro-4-methylphenyl,3-chloro-4-fluorophenyl, 3,4-difluorophenyl, 3-trifluoromethylphenyl,3-trifluoromethyl-4-chlorophenyl, 3-methoxyphenyl, 3-methylphenyl,3-ethylphenyl, and 3-isopropylphenyl; or R¹ is chosen from2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl,2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl,2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl,2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 2-chlorophenyl,2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl,2,6-dichlorophenyl, 2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl,2,3,6-trichlorophenyl, 2,4,5-trichlorophenyl, and 2,4,6-trichlorophenyl;or R¹ is chosen from 2-methylphenyl, 4-methylphenyl, 2,3-dimethylphenyl,2,4-dimethylphenyl, 2,5-dimethyl-phenyl, 2,6-dimethylphenyl,3,4-dimethyl-phenyl, 2,3,4-trimethylphenyl, 2,3,5-trimethyl-phenyl,2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl,2-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl, 2,4-diethylphenyl,2,5-diethylphenyl, 2,6-diethylphenyl, 3,4-diethylphenyl,2,3,4-triethylphenyl, 2,3,5-triethylphenyl, 2,3,6-triethylphenyl,2,4,5-triethylphenyl, and 2,4,6-triethylphenyl; or R¹ is chosen from2-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl,2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl, 2,6-dimethoxyphenyl,3,4-dimethoxyphenyl, 2,3,4-trimethoxyphenyl, 2,3,5-trimethoxyphenyl,2,3,6-trimethoxy-phenyl, 2,4,5-trimethoxyphenyl, 2,4,6-trimethoxyphenyl,2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2,3-dihydroxyphenyl,2,4-dihydroxyphenyl, 2,5-dihydroxyphenyl, 2,6-dihydroxyphenyl,3,4-dihydroxy-phenyl, 2,3,4-trihydroxyphenyl, 2,3,5-trihydroxy-phenyl,2,3,6-trihydroxyphenyl, 2,4,5-trihydroxyphenyl, and2,4,6-trihydroxyphenyl.
 2. A compound according to claim 1 wherein R²and R³ are each independently chosen from hydrogen, methyl, ethyl,n-propyl, iso-propyl, cyclopropyl, n-butyl, iso-butyl, sec-butyl, andtert-butyl; R is a unit chosen from —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃),—N(CH₂CH₃)₂, —N(CH₃)(CH₂CH₃), —NH(CH₂CH₂CH₃), —N(CH₃)(CH₂CH₂CH₃),—N(CH₂CH₃)(CH₂CH₂CH₃), and —N(CH₂CH₂CH₃)₂; and wherein R¹ is chosen from2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl,2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl,2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl,2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 2-chlorophenyl,2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl,2,6-dichlorophenyl, 2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl,2,3,6-trichlorophenyl, 2,4,5-trichlorophenyl, and 2,4,6-trichlorophenyl.3. A compound according to claim 1 wherein R² and R³ are eachindependently chosen from hydrogen, methyl, ethyl, n-propyl, iso-propyl,cyclopropyl, n-butyl, iso-butyl, sec-butyl, and tert-butyl; R is a unitchosen from —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃), —N(CH₂CH₃)₂,—N(CH₃)(CH₂CH₃), —NH(CH₂CH₂CH₃), —N(CH₃)(CH₂CH₂CH₃),—N(CH₂CH₃)(CH₂CH₂CH₃), and —N(CH₂CH₂CH₃₎ ₂; and wherein R¹ is chosenfrom 2-methylphenyl, 4-methylphenyl, 2,3-dimethylphenyl,2,4-dimethylphenyl, 2,5-dimethyl-phenyl, 2,6-dimethylphenyl,3,4-dimethyl-phenyl, 2,3,4-trimethylphenyl, 2,3,5-trimethyl-phenyl,2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl,2-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl, 2,4-diethylphenyl,2,5-diethylphenyl, 2,6-diethylphenyl, 3,4-diethylphenyl,2,3,4-triethylphenyl, 2,3,5-triethylphenyl, 2,3,6-triethylphenyl,2,4,5-triethylphenyl, and 2,4,6-triethylphenyl.
 4. A compound accordingto claim 1 wherein R² and R³ are each independently chosen fromhydrogen, methyl, ethyl, n-propyl, iso-propyl, cyclopropyl, n-butyl,iso-butyl, sec-butyl, and tert-butyl; R is a unit chosen from —NH₂,—NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃), —N(CH₂CH₃)₂, —N(CH₃)(CH₂CH₃),—NH(CH₂CH₂CH₃), —N(CH₃)(CH₂CH₂CH₃), —N(CH₂CH₃)(CH₂CH₂CH₃), and—N(CH₂CH₂CH₃)₂; and wherein R¹ is chosen from 2-methoxyphenyl,4-methoxyphenyl, 2,3-dimethoxyphenyl, 2,4-dimethoxyphenyl,2,5-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,4-dimethoxyphenyl,2,3,4-trimethoxyphenyl, 2,3,5-trimethoxyphenyl, 2,3,6-trimethoxy-phenyl,2,4,5-trimethoxyphenyl, 2,4,6-trimethoxyphenyl, 2-hydroxyphenyl,3-hydroxyphenyl, 4-hydroxyphenyl, 2,3-dihydroxyphenyl,2,4-dihydroxyphenyl, 2,5-dihydroxyphenyl, 2,6-dihydroxyphenyl,3,4-dihydroxy-phenyl, 2,3,4-trihydroxyphenyl, 2,3,5-trihydroxy-phenyl,2,3,6-trihydroxyphenyl, 2,4,5-trihydroxyphenyl, and2,4,6-trihydroxyphenyl.
 5. A compound having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄linear or branched alkyl; and R⁵ is chosen from: i) C₁-C₄ linear,branched, or cyclic alkyl; ii) halogen; iii) —OR⁶; iv) —SO₂N(R⁶)₂; v)—CH_(m)X_(n); wherein each X is independently F, Cl, Br, or I, m is from0 to 2, m+n =3; vi) —NO₂; and vii) phenyl; each R⁶ is independentlyhydrogen, or C₁-C₄ linear, branched, or cyclic alkyl; excludingN2-(4-chlorophenyl)-N⁴-[(3-diethylamino)propyl]-pyrimidine-2,4-diamine.6. A compound according to claim 5 wherein R² and R³ are eachindependently hydrogen, methyl, and ethyl.
 7. A compound according toclaim 5 wherein R⁵ is chosen from 3-chloro, 4-chloro, 3,4-dichloro,3-chloro-4-methyl-, 3-chloro-4-fluoro, 3,4-difluoro, 3-trifluoro-methyl,3-trifluoromethyl-4-chloro, 3-methoxy, 3-methyl, 3-ethyl, and3-iso-propyl.
 8. A compound according to claim 5 wherein R⁵ is chosenfrom 2-fluoro, 3-fluoro, 4-fluoro, 2,3-difluoro, 2,4-difluoro,2,5-difluoro, 2,6-difluoro, 2-chloro, 2,3-dichloro, 2,4-dichloro,2,5-dichloro and, 2,6-dichloro.
 9. A compound according to claim 5wherein R⁵ is chosen from 2-methoxy, 4-methoxy, 2,3-dimethoxy,2,4-dimethoxy, 2,5-dimethoxy, 2,6-dimethoxy, 3,4-dimethoxy, 2-hydroxy,3-hydroxy, 4-hydroxy, 2,3-dihydroxy, 2,4-dihydroxy, 2,5-dihydroxy,2,6-dihydroxy, and 3,4-dihydroxy.
 10. A compound having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄linear or branched alkyl; and R⁵ is chosen from: i) C₁-C₄ linear,branched, or cyclic alkyl; ii) halogen; iii) —OR⁶; iv) —SO₂N(R⁶)₂; v)—CH_(m)X_(n); wherein each X is independently F, Cl, Br, or I, m is from0 to 2, m+n =3; vi) —NO₂; and vii) phenyl; each R⁶ is independentlyhydrogen, or C₁-C₄ linear, branched, or cyclic alkyl; and L is chosenfrom: i) —CH₂—, methylene; ii) —CH₂CH₂—, ethylene; iii) —CH(CH₃)CH₂—,1-propylene; iv) —CH₂CH(CH₃)—, 2-propylene; v) —CH₂CH₂CH₂CH₂—, butylene.vi) —CH(CH₃)CH₂CH₂—, 1-butylene; vii) —CH₂CH(CH₃)CH₂—, 2-butylene; andviii) —CH₂C(CH₃)₂CH₂—, 2,2-dimethylpropylene; excludingN²-(4-chlorophenyl)—N⁴-[(2-diethylamino)ethyl]-pyrimidine-2,4-diamineandN²-(4-methylphenyl)—N⁴-[(2-diethylamino)ethyl]-pyrimidine-2,4-diamine.11. A composition comprising: A) the compound of claim 1 and B) thebalance carriers and excipients.